GENOMICS8,525-535(1990)

A Molecular

Genetic Linkage Map of Mouse Chromosome ANN

Department

of Medicine,

7

M. SAUNDERS AND MICHAEL F. SELDIN Duke University Medical Received

March

28, 1990;

Center, Durham, North Carolina 27710

revised

July 17, 1990

A rapid and efficient tool in the delineation of gene linkage relationships in the mouse genome is provided by using interspecific crosses (reviewed by Avner et al., 1988). The mapping of molecular markers requires identifying a restriction fragment length variant (RFLV) between the two strains used in the segregation analysis. While intraspecific crosses and recombinant inbred strain analyses have been useful in mapping many phenotypic, molecular, and biochemical markers, it is often difficult to find RFLVs for cloned genes because most inbred mouse strains are closely related (Ferris et al., 1982). The strength of interspecific backcross analysis resides in the evolutionary distance between the two parental species often used in interspecific backcross analyses, Mus domesticus (represented by C3H/HeJ-gld/gld in this study) and Mu.s spretus. The divergence of these two species approximately 3-5 million years ago (Bonhomme et al., 1984) has allowed for the accumulation of DNA sequence differences; consequently, RFLVs are easily detected, and large numbers of loci can be mapped at the same time using a single backcross DNA panel (reviewed by Avner et al., 1988). In an earlier study, we reported on the linkage relationships of 16 loci on mouse chromosome 7 and the homology between proximal mouse chromosome 7 and a region of human chromosome 19 that contains the myotonic dystrophy (DM) locus (Saunders and Seldin, 1990). The current study was undertaken to extend the linkage relationships of molecular probes on mouse chromosome 7. Interspecific backcross analysis was used to define RFLVs for 14 additional probes (Rip, Tgfb, Gpi, Cebp, Rras, Kal, Egfbp, Emv-23, D7Was12, Pth, Hbb, H-19, Th, and Fis-1) that have been assigned to mouse chromosome 7 by either limited genetic analyses using somatic cell hybrid panels or recombinant inbred strains and backcrosses (Davisson et al., 1989; Lalley et al., 1989; Lyon, 1989). The segregation of Cyp2a, D19Fl lSlh, Myod-1, Otf-2, Rnu2pa, Rnulp70, and Xrcc-1, previously unmapped in the mouse, was also examined. We have examined the positions of these 21 loci

The homology between mouse chromosome 7 and human chromosomes 11,15, and 19 was examined using interspecific backcross animals derived from mating CSH/HeJ-gZd/ gld and Mus spretua mice. In an earlier study, we reported on the linkage relationships of 16 loci on mouse chromosome 7 and the homologous relationship between this chromosome and the myotonic dystrophy gene region on human chromosome 19. Segregation analyses were used to extend the gene linkage relationships on mouse chromosome 7 by an additional 21 loci. Seven of these genes (Cyp2a, DlSFllSlh, Myod-l, Otf-2, Rnulp70, Rnu2pa, andxrccI) were previously unmapped in the mouse. Several potential mouse chromosome 7 genes (Mel, Hkr-1, Icam-1, Pus) did not segregate with chromosome 7 markers, and provisional chromosomal assignments were made. This study establishes a detailed molecular genetic linkage map of mouse chromosome 7 that will be useful as a framework for determining linkage relationships of additional molecular markers and for identifying homologous disease genes in mice and humans. 0 1000 Academic Press, Inc.

INTRODUCIION

A major impetus in constructing genetic linkage maps of human chromosomes is the localization of genes involved in inherited disorders. By determining what chromosomal segments are conserved in the mouse and human genomes, it will be possible to make predictions about where mutations localized in one species will map in the other (reviewed by Nadeau, 1989). For an inherited human defect, mapping of the homologous mouse region could facilitate the isolation of a disease gene by indicating the probable location of its mouse homolog and by identifying closely linked markers. Comparative mapping is also useful for developing mouse model systems to study human genetic diseases. The utility of this approach was demonstrated by the establishment of a relationship between the mouse muscular dystrophy (mdx) mutation and the human Duchenne muscular dystrophy locus (Ryder-Cook et aZ., 1988). 525

All

Copyright 0 1990 rights of reproduction

osss-7543/90 $3.00 by Academic Press, Inc. in any form reserved.

526

SAUNDERS

relative to our laboratory’s earlier map using the same panel of interspecific backcross progeny and have established a 68.7-CM linkage map of mouse chromosome 7. MATERIALS

AND

METHODS

Mice C3H/HeJ-gM/gld (generalized lymphoproliferative disease), M. spretus (Spain), and [(C3H/HeJ-gkl/gZd X M. spretus)F, X C3H/HeJ-gti/gld] backcross mice were bred and maintained as described earlier (Seldin et aZ., 1988). Southern Hybridization Genomic DNA was obtained from C3H/HeJ-gZd/ gld, M. spretus, and [(C3H/HeJ-gld/gld X M. spretus)F, x C3H/HeJ-gZd/g2d] mice by standard techniques. These samples were used to examine the segregation of RFLVs detected with molecular probes. Ten-microgram samples were digested with restriction endonucleases (Boehringer-Mannheim Biochemicals, Indianapolis, IN) and electrophoresed in 0.9% agarose gels. DNA was transferred to Nytran membranes (Schleicher & Schuell, Inc., Keene, NH) and hybridized at 65°C as described previously (Seldin et al., 1988). The Nytran membranes were then washed at either 65°C at 0.2X SSC final (high) stringency or 58°C at 0.5X SSC final (low) stringency. Molecular Probes All probes were labeled by the hexanucleotide technique with [Ly-32P]dCTP (3000 Ci/mmol; New England Nuclear, Boston, MA) using the oligolabeling kit and protocol from Pharmacia Fine Chemicals. Probes used in this study (Table 1) include the human APOCl cDNA clone pUCI-A4 (Knott et al., 1984), the human APOC2 cDNA clone pKT218 (Humpheries et aZ., 1984), the mouse Apoe cDNA clone p2CL (Lusis et al., 1987), the human APS cDNA clone phPA-S (Sutherland et al., 1988), the rat cDNA Atpa- clone pA1 (Kent et al., 1987), the human BCL3 genomic clone pcul.4~ (McKeithan et al., 1987; Korneluk et al., 1989), an EcoRI-linearized plasmid containing a rat cDNA Calc clone (clone not designated, Jacobs et al., 1981), the human CEA cDNA clone pCEA2 (Zimmerman et al., 1987), the mouse Cebp cDNA clone Cebp I (Xanthopoulos et al., 1989), the human CGB5 and CGB6 genomic clones P-hCG5 and @-hCG6 (Talmadge et al., 1983), the human CKMM cDNA clone pJN2CK-M (Nigro et al., 1987), the human CYP2A cDNA clone P450 IIA3 (Yamano et aZ., 1989), the rat Cyp2b cDNA clone pP-450-5 (Simmons et aZ., 1985), the mouse D7Rp2 cDNA clone pMK908 (Berger et al.,

AND

SELDIN

1981), a mouse D7Was12 genomic clone (clone not designated, Disteche and Adler, 1984), the human Dl9FllSl cDNA clone OL5 (Jandel et al., 1985), the human D19S7 genomic clone ~4.1 (Shaw et al., 1986), the human D19S8 genomic clone ~17.1 (Shaw et al., 1986), the human Dl9S9 genomic clone pIJ2 (Shaw et al., 1986), p17.4-4(2), a cDNA probe detected with the anonymous human chromosome 19 clone LDR152 (Dl9Sl9, Bartlett et al., 1987; J. Gilbert, personal communication), the human D19S51 genomic clone p134C (K. Johnson, St. Mary’s Hospital, London, personal communication), the mouse Egfbp cDNA clone MB2-12A (Blaber et al., 1987), the mouse Emu23 genomic clone pEmv-23 (probe 23.3, Rinchik et aZ., 1989), the human ERCCl cDNA clone pE12-12 (van Duin et al., 1986), the human ERCC2 genomic clone pKER2 (Weber et al., 1988), a l.O-kb BgZII fragment of the human oncogene FES clone ~80 (Sodroski et al., 1984), the mouse Fis-1 genomic clone ~1.8 (Silver and Buckler, 1986), the human GPI cDNA clone HS10 (GenBank HUM-NLK KO 3515, May 1987; M. Gurney, Northwestern University, personal communication), the human HBB genomic clone pBR$ a (US Patent 4,683,194; Lawn et al., 1978), the human HKRl genomic clone pHKR1 RS-RsaI-680 and the human HKR2 genomic clone pHKR2 HH-HinfI -325 (Ruppert et al., 1988), a 0.9-kb Hind111 fragment of the human HRASl clone HB-11 (Ellis et al., 1981), the mouse H-19 cDNA clone H19RS (Pachnis et al., 1984), a mouse Icam-l cDNA clone (A. Brian and P. Kuhlman, University of California, San Diego, personal communication), the human IGFlR cDNA clone pIGF-l-R.8 (Ullrich et al., 1986), the human IGF2 cDNA clone pIGF2/8-1 (Dull et al., 1984), the human INS genomic clone phins 214 (Bell et al., 1984), the mouse KuZ cDNA clone pMK-1 (Richards et al., 1982), the human LHB genomic clone /3 LH (Talmadge et al., 1983), the rat Mag cDNA clone 5Mag (Barton et al., 1987), the human MEL genomic clone PEmbl 600 (Padua et al., 1984), the human MYODl cDNA clone pV2Clla (Davis et aZ., 1987), the mouse Ngfg partial cDNA clone pSM676 (Howles et al., 1984), the human OTF2 partial cDNA clone Put-3-l (Ko et aZ., 1988), the human PRKCG cDNA clone phPKC-y 6 (Coussens et al., 1986), a 450-bp NcoI-XbaI fragment of the rat Pth genomic clone rPTHs-1 (Heinrich et aZ., 1984; Lalley et al., 1987), the mouse Rip cDNA clone pf3 (Noshiro et al., 1988), the mouse Rras cDNA clone MR-9 (Lowe et al., 1987), the mouse Sau-3 cDNA clone pRS48 (Stearman et al., 1986), the mouse Tgfb cDNA clone pMur/32 (Derynck et al., 1986), the human U2 snRNP-specific A’ polypeptide cDNA clone A’910 (Fresco et al., 1987; Fresco and Keene, manuscript in preparation), the human SNRP70 cDNA clone FL70K (Query et al., 1987), the human TH genomic clone pHGTH4 (O’Malley and Rotwein, 1988), the mouse Tyr cDNA clone

LINKAGE

MAP

OF

MOUSE

CHROMOSOME

527

7

MTYSllC

(Kwon et aZ., 1987), and the human cDNA clone pXPl-30 (Sicilian0 et al., 1986). Nytran membranes hybridized with probes for the following loci were washed under low stringency conditions: APOCI, APOCZ, APS, BCL3, CEA, D19S7,

CypZb, Rip, Tgfb]-2.7

XRCCl

CM-[Gpi, Mag]. Assuming this gene order, there were no double crossovers in 334 meiotic events within this 8.1-cM interval.

D19S8, D19S9, D19S51, GPI, HBB, HKRl, HKRZ, MEL, MYODl, and TH. Membranes hybridized with

Segregation Analysis Chromosome 7

the remaining probes gency conditions.

Our previous study included eight loci (D7Rp2, Lhb, Ngfg, Hras-1, Igflr, Fes, Tyr, and Calc) that were

Statistical

were washed

under high strin-

Methods

Gene linkage was determined by segregation analysis (Green, 1981). Gene order was established by minimizing the number of double recombinants between genes that were determined to be within a linkage group and confirmed by maximum likelihood analysis (Bishop, 1985). The deviation of recombination frequency from a Poisson distribution was calculated using a x2 test with Yates correction factor (Daniel, 1987). RESULTS

Identification of Informative Each Molecular Probe

Restriction

Bands for

Each molecular probe was hybridized to Southern blots containing DNA from the homozygous parent generation C3H/HeJ-gld/gld (CC) and the heterozygous F, generation (C3H/HeJ-gZd/gld X M. spretus) (SC). With each probe, at least one endonuclease-digested DNA panel displaying unique M. spretus RFLVs was identified (Fig. 1; and Saunders and Seldin, 1990). The restriction endonucleases used were EcoRI, BamHI, BglII, TaqI, MspI, PvuII, and P&I. To examine the segregation of RFLVs, subsequent digestions were performed on DNA samples obtained from [(C3H/HeJ-gld/gld X M. spretus)F, x C3H/ HeJ-gld/gld] interspecific backcross progeny using the appropriate endonucleases. RFLVs from individual backcross mice exhibited either the CC or the SC pattern for each locus examined.

Segregation Analysis Chromosome 7

of 15 Loci on Proximal

Mouse

Segregation analysis of CypZa, Rip, Tgfb, Xrcc-1, Otf-2, D19Fl IS1 h, and Gpi extended our earlier examination of gene linkage relationships on proximal mouse chromosome 7 that included Pkcg, Apoe, Atpa2, Ckmm, D19S19h, Ercc-2, CypZb, and Mag. These genes were typed for RFLVs in 334 backcross mice. Minimization of crossover frequency of these loci (Fig. 2) resulted in the following gene order (+ standard deviation): (centromere)-Pkcg-1.5 f 0.7 CM-

[Apoe, Atpa-2,

Ckmm,

D19S19h,

CM-Xrcc-I-0.6

f 0.4 CM-Otf-2-0.9

Ercc-21-0.3

f 0.3

f 0.5 CM-[CypZu,

-t 0.9 CM-DlSFll

Sl h-2.1 + 0.8

of 22 Loci on Distal

Mouse

mapped to a 40-CM segment of distal mouse chromosome 7 (Saunders and Seldin, 1990). In the current study, an additional 14 loci were localized within a 61-CM interval telomeric to Mug/Gpi by segregation analysis of RFLVs determined in 114 backcross mice DNA samples. The human homologs of these loci, when known, are distributed among chromosomes 11, 15, and 19. Minimization of crossover frequency (Fig. 2) resulted in the following gene order (+ standard deviation): (centromere)-[Pkcg-Mag/Gpi]-0.9 f 0.9

CM-[D7RpZ/Cebp]-10.5 it 2.9 CM-[Egfbp, Kal, Lhb, Ngfg, Rnulp70, Rras]-0.9 f 0.9 CM-Myod-l-9.6 f 2.8 CM-RnuZpa-1.8 -+ 1.2 CM-[Hras-1, Igflr]-6.1 + 2.4 CM-Fes-4.4 +- 1.9 CM-Tyr-0.9 + 0.9 CM-Emu-23-0.9 f 0.9 CM-D7Wus12-7.0 f 2.4 CM-[C&c, Pth]-0.9 f 0.9 CM-Hbb-14.9 f 3.3 CM-H-19-0.9 f 0.9 CM-Th0.9 * 0.9 CM-Fis-1. There were 14 double crossovers in 114 meiotic events within the 68.7-CM interval between Pkcg and Fis-1; no triple or greater crossovers were observed. Over the entire segment of 68.7 CM, the number of double crossovers observed did not differ significantly (P > 0.05) from the number expected. However, a decrease in double crossovers, a phenomenon known as positive interference, was seen in the 51-CM interval between Pkcg and Hbb. If recombination events were distributed randomly over the chromosome (i.e., if there was no interference), chromosomes with zero, one, and two or more recombinations within this 51CM segment would follow a Poisson distribution with a mean of 0.51 (58 crossovers/l14 meiotic events). The number of crossovers expected would be 68.4, 34.9, and 8.0; however, 59, 52, and 3 were observed. This deviation is statistically significant: x2 = 12.77 (1 df) P < 0.0005). The lower than expected number of double crossovers in this 51-CM interval is consistent with prior observations in both interspecific (Ceci et al., 1989; Kingsley et al., 1989; Moseley and Seldin, 1989; Seldin et al., 1989; Siracusa et al., 1989) and laboratory strain (Blank et al., 1988) backcross studies.

Examination of Other Potential 7 Genetic Markers

Mouse Chromosome

Fifty-four probes were initially screened for crosshybridization with mouse genomic DNA, unique M. spretus RFLVs, and segregation with other mouse

528 a

SAUNDERS Cebp CC SC

Cyp20 CC SC

D7Was12

DI9Fl?Slh

CC SC

SELDIN

Emv-23

4 fbp CC SC

cc SC

AND

cc SC

b rns CC SC

Kal

Me/

cc SC

cc SC

O/f-Z , cc SC

Pth

Pvs

cc SC

CC SC -23.1 -9.4 -6.6

l -

-

-1.1

-

BqmHI

ECORI

ECCf?I

-2.3 -2.0

1.3

-1.3

EcoRI

- 1.1

BamHI

Taq I

Taq I ITS-I

cc SC

Gpi cc SC

H-19

Hbb

cc K

cc SC

Hkr-

I

TWI

BamHI

Rnu2po

RlWIp7V

PstI

hIspI

hd?I

Icam-

ccsc

cc SC

Rip -23.1

cc SC

cc SC

cc SC

Rras

Tgfb

Th

cc SC

cc SC

cc Y

EcoRI

P5f I

ToqI

xrcc-7 cc SC

-9.4

l

-6.6

-4.3

-2.3 -2.0

ä E,+gyg

_

a%-

MSPI

M.91

BglIl

-

J MspI

EcoRI

*

-1.3 -1.1

EcoRI

EcoRI

MspI

Taq I

&ImHI

FIG. 1. Southern blot identification of unique Mus spretus RFLVs detected molecular probes. Arrowheads signify bands present in DNA from (CBH/HeJ-gld/gld X Mus spretus) F, (SC) not present in homozygous C3H/HeJ-gld/gld (CC) mice. Where multiple restriction bands are observed (Z&d, Rip, Egfbp), they segregated together. The informative restriction endonucleases are shown below each panel, and the molecular weight standards (in kb) are shown on the right. RFLV information on probes not shown was previously reported (67).

chromosome 7 genetic markers. Of the 22 human chromosome 19-specific probes screened in this study, 10 of them did not cross-hybridize with mouse genomic DNA under low stringency wash conditions. These included the probes for the following loci:

APOCl, APOC2, APS, BCL3, CEA, D19S7, Dl9S8, D19S9, D19S51, and HKR2. A second problem encountered was the hybridization of some probes to repetitive elements in mouse genomic DNA. Restriction endonuclease digestion of the full-length clones for Fes, Hras-1, and Pth resulted in smaller fragments that did not hybridize to mouse repetitive elements and recognized unique M. spretus variants. Informative fragments of the clones for CGB5 and CGB6 were not found. Five clones detected RFLVs that did not segregate with the mouse chromosome 7 markers mapped in this study. The selection of these clones was based on their earlier assignments to either mouse chromosome 7 or human chromosomes 19 and 11. The mouse homolog of MEL was mapped to mouse chromosome

8 which shares homology with human chromosome with the 19p13 (Ceci et aZ., 1990). This is consistent recent reassignment of MEL from human chromosome 19p13.2-q13.2 (Shaw and Eiberg, 1987) to human chromosome 19p13.2-ten (Nimmo et aZ., 1989). and PVS cosegreThe mouse homologs of ICAMl gated with markers on mouse chromosome 9. PVS has been regionally assigned to human chromosome 19q12-q13.2 (Siddique et al., 1988), and ICAMl has been placed on human chromosome 19 (Greve et al., 1989). Of note, mouse chromosome 9 also shares homology with human chromosome 19p13 (Kingsley et al., 1989). We have tentatively localized the mouse homolog of HKRl to mouse chromosome 17 which contains a linkage group homologous to human chromosome 19q12 (Nadeau, 1989). HKRl was recently mapped to human chromosome 19q by somatic cell hybrid panels (Ruppert et al., 1988). The unique M. spretus variants detected by the human probe for INS, assigned to human chromosome 11~15.5 (Junien and McBride, 1989), did not segregate with

LINKAGE

MAP

urn321232734 307 5 123

6

Pkcg

ClusterI+ Xrcc- 1 Otf-2 ClusterIP DlSFllSlh

9

OF

MOUSE

CHROMOSOME

7

529

7

;’

1;

,, j, t

Gpi,&fag

;i D?Rp2, Cebp ;; Cluster lU* ‘_; Myad-l :i Rnu2pa Was- 1, Igflr :: Fas $1 $I TY~ Emv-23 $ D7Was12 L; ?‘,’ Co/c, Pth Hbb r’. H-19 .:i Jj Th jr Fis-1 25

7.

1116115 -----111

7

1

i

FIG. 2. Analyses of interspecific backcross haplotypes. (A) Haplotype distribution for mouse chromosome 7 loci located within the syntenic human chromosome 19 segment that contains DM. For these loci, 334 backcross progeny were examined. (B) Haplotype distribution for the 37 loci mapped to mouse chromosome 7. These data include 114 of the 334 backcross progeny shown in A. Each column represents a chromosomal haplotype identified in the backcross progeny that was inherited from the (CBH/HeJ-gld/gtd x M. spretus)F, parent. The number of backcross mice with each observed haplotype is indicated at the bottom of each column. Solid bars represent M. spretus-like RFLVs, and hatched bars represent CBH/HeJ-gZd/gWlike RFLVs as determined by Southern blot hybridization with genetic probes (Fig. 1). The Cluster* designation includes (I) Apoe, Atpa-2, Ckmm, D19Sl9k, and Ercc-2 in which no crossovers were observed in 334 backcross progeny; (II) Cyp2a, CypZb, Rip, and Tgfb in which no crossovers were observed in 334 backcross progeny; and (III) Egfbp, Kal, Lhb, Ngfg,Rnulp70, and Rras in which no crossovers were observed in 114 backcross progeny.

mouse chromosome 7 markers. This locus, designated as Ins-2, has been mapped to mouse chromosome 7 by somatic cell hybrid panels (Lalley and Chirgwin, 1984). Ongoing studies may determine if the unique variants recognized by the human INS clone are segregating with either of the putative mouse insulin genes found on mouse chromosomes 6 and 15 (Lalley and Chirgwin, 1984). Unique M. spretus variants were not found for the ERCCl, IGF2, and Soa- clones used in this study. The enzymes used were EcoRI, BanHI, BglII, To@, MspI, PuuII, and PstI.

DISCUSSION

We have established a multilocus linkage map of mouse chromosome 7 that includes 37 genetic markers (Fig. 3). Marker density in this study was 0.31 loci/CM (21 different loci mapped over a 68.7-CM distance, with the largest distance between two markers being 14.9 CM). This interspecific backcross linkage map will be useful as a framework for determining linkage relationships of additional cloned

DNA markers and for identifying homologous disease genes in humans and mice. Comparison with Previous Mapping Data An important consideration in interspecific backcross mapping studies is whether the data are consistent with results obtained by intraspecific crossesand recombinant inbred strain analyses, as well as by cytogenetic and somatic cell hybrid studies. Our map does not differ significantly from that of Lyon (1989), which is a computer-generated map of all published data from two- and three-point crosses. As a result, both genetic distances and order of loci in this composite map are very approximate. With the exception of an inverted order between Th and Fis-1, which are closely linked on both maps, the relative regional localizations of genes are very similar. The interval between Atpa-2, the most centromeric marker on both maps, and Fis-l/Th, the most telomeric markers on both maps, is nearly the same. The major difference between the two maps is in the genetic distances between loci. This may be a result of comparing a map established in a single panel of backcross progeny

530

SAUNDERS

AND

SELDIN

[D7Rp2,

Pkcg Apoe,Atpa-2,

Ckmm,

--

3.3 6.0 6.1 9.0

-----

Cyp2aa,Cyp2b,Rip, 019FllSlh

Jgfb

Gpf,Mag 07R@,Cebp

19.5

--

Egfbp,Kal,Lhb,fVgfg,Rnulp70,

20.4

--

Myod-1

30.0 31.8

---

Rnu2pa

37.9

--

Fes

42.3 43.2

----

Jyr Emv-23 D7Was12

51.1 52.0

--

Catc,Pth

--

Hbb

66.9

----

H-19 Jh Fis-1

44.1

67.8 68.7

Rras

Homology

Ngfg,

Rras,

with Human

Chromosomes

11 and 15

The linkage relationship of 12 additional loci that have homologs, when known, on human chromosomes 11 and 15 (Table 1) was also examined. These genes were localized within a 49.2-cM interval telomerit to the [Egfhp * . . Rnulp70] gene cluster. Their order is (centromere)-(Pkcg-[Egfbp. . . Rnulp70])-

Myod-1-Rnu2pa-[Hras-1, Igflr]-Fes-Try-Emv-23D7Wasl2-[Calc, Pth]-Hbb-H-19-Th-Fis-l-(telo-

of mouse chromosome 7. (CM) for mouse chromoof crossover events using Table 1.

with a map compiled from numerous studies variety of crosses and different anchor loci.

with Human

Lhb,

in both species, occurred during the evolution of the two species. However, the gene order appears to be the same in both species within the Pkcg/PRKCG-Mag/ MAG chromosomal segment.

Hras-l,/gflr

FIG. 3. Molecular genetic linkage map Gene placement and mapping distances some 7 were determined by minimization the recombination frequencies shown in

Homology

Kal,

DlSFllSl-[CYP2A, CYP2B, TGFBl]-[XRCCl, ATPlA3] - D19S19 - APOE - CKMM - ERCC2 [PRKCG, RRAS, LHB, KLKl, SNRP70]-(telomere) (LeBeau et al., 1989). The data suggest that an inversion extending from Pkcg through the interval between [Mag, Gpi] and [Egfbp, Kal, Lhb, Ngfg, Rras, Rnulp70], a cluster of genes which remains telomeric

D 19.5 79h, Ercc-2

Xrcc-1 Ott-2

1.0 2.4

--

Cebp]-[Egfbp,

Rnulp70]-(telomere). Human physical and genetic linkage studies within the syntenic 19q13.1-q13.4 region indicate an order of (centromere)-GPI-MAG-

Chromosome

using a

19q

A 19.5-cM molecular genetic map of the proximal segment of mouse chromosome 7 was derived using segregation analyses from 334 meiotic events for 15 probes and 114 meiotic events for six others. All of these loci are found on human chromosome 19q13.113.4 with the exception of the five genes that have not been mapped in humans (Table 1). As noted in our previous study (Saunders and Seldin, 1990), comparison of mouse and human gene orders indicates that gene orientation with respect to the centromere has not remained intact within the syntenic group. The order of genes on proximal mouse chromosome 7 from the centromere is Pkcg-[Apoe,

Atpa-2, Ckmm, D19S19h, Ercc-2]-Xrcc-I-Otf-2[Cyp2a, Cyp2b, Rip, Tgfb]-DlgFllSlh-[Mag, Gpi]-

mere). As noted in our earlier study, the interval between Igflr and Fes represents a 6.1-CM gene linkage group that is homologous with human chromosome 15q25-q26 (Saunders and Seldin, 1990). Since there were no recombination events between Igflr and Hras-1, a locus that maps to human chromosome 11~15.5, the immediate area around Igflr may represent the proximal border of the human 15q homologous linkage group. Mouse chromosome 7 has a number of genes with homologs found on both the p- and q-arms of human chromosome 11; however, gene order does not appear to be conserved and is interrupted by the human chromosome 15 linkage group (Fig. 3). The human homologs of these genes are clustered at the ends of the p- and q-arms of chromosome 11 (Junien and McBride, 1989). The extent of conservation between human and mouse genes located near the tips of chromosomes should become more apparent as more detailed genetic linkage maps are generated. The mouse X chromosome is another example of a difference between human and mouse chromosome morphology and conserved linkage group order (Yang-Feng et al., 1986). It has been proposed that a small number of ancestral intrachromosomal rearrangements would be sufficient to account for these differences (Buckle et al., 1985; Searle et al., 1987).

Clusters of Genes on Mouse Chromosome

7

The presence of gene clusters indicates either close physical proximity or small inversions or deletions in M. spretus chromosomes relative to C3H/HeJ-gld/

LINKAGE

MAP

OF

MOUSE

TABLE

CHROMOSOME

531

7

1 95% Confidence limits

Gene

name

Protein kinase c, y Apolipoprotein E Na,K-ATPase, a Creatine kinase M, muscle form DNA segment, single copy, probe LDR152 Excision repair 2 X-ray repair 1 Octamer-specific DNA binding protein 2 Cytochrome P450 IIA3 Cytochrome P450, 2b, phenobarbitol inducible Regulator of phenobarbitol inducible P450 expression Transforming growth factor, fl DNA segment, numerous copies, probe OL5 Myelin-associated glycoprotein Glucose phosphate isomerase DNA segment, Roswell Park 2 DNA binding protein C/EBP Luteinizing hormone, fl Nerve growth factor, y Epidermal growth factor binding protein Kallikrein Harvey rat sarcoma oncogene, subgroup R Ul small ribonucleoprotein 70kDa polypeptide Myogenic differentiation 1 U2 small ribonucleoprotein polypeptide A Harvey rat sarcoma oncogene Insulin-like growth factor I receptor Feline sarcoma oncogene Tyrosinase Ecotropic MuLV-23 DNA segment, Washington 12 Calcitonin Parathyroid hormone Hemoglobin, @ chain H-19 mRNA Tyrosine hydroxylase Friend virus integration site-l

Locus

Linkage

Number of recombinations

interval

r(cM)

Low

High

Human

homolog”

Pkcg Apoe AtpaCkmm

Pkcg-Apoe Apoe-AtpaAtpa-2-Ckmm Ckmm-DlSSlSh

5/334 o/334 o/334 o/334

1.5

0.04

0.0 0.0 0.0

0.00 0.00 0.00

1.76 1.02 1.02 1.02

PRKCG APOE ATPlA3 CKMM

lSq13.4 19q13.2 19q12-q13.2 lSq13.2-q13.3

Dl9S19h Ercc-2 Xrcc-1

DlSSlSh-Exe-2 Ercc-P-Xrcc-1 Xrcc-I-Otf-2

o/334 l/334 2/334

0.0 0.3 0.6

0.00 0.00 0.00

1.02 1.17 1.33

Dl9Sl9 ERCC2 XRCCl

lSqcen-q13.2 19q13.2-q13.3 lSq13.1-q13.2

otf-2

Otf-2-Cyp2a Cyp2a-Cyp2b

31334 o/334

0.9

0.01

CypZa

0.0

0.00

1.54 1.02

OTF2 CPY2A

1% lSq13.1-q13.2

Cw2b

Cyp2b-Rip

o/334

0.0

0.00

1.02

CYP2B

lSq13.1-q13.2

Rip Tgfb

Rip-Tgfb Tgfb-DlSFllSlh

o/334 g/334

0.0 2.7

0.00 0.18

1.02 2.42

N.D.b

DlSFllSlh

2.1

0.07

0.0

0.00

Gpi D7Rp2 Cebp Lhb Ngfg

DlSFllSlh-Mag Mag-Gpi Gpi-D7Rp2 D7Rp2-Cebp Cebp-Lhb Lhb-Ngfg Ngfg-Egfbp

1.99 1.02 4.62 3.07 15.55

Egfbp Kal

Egfbp-Kal Kal-Rras

Rras

RrasRnulp70

Rnulp7(P Myod-1

Rnulp70-Myod-1 Myod-I-Rnu2pa

Rnu2pa’ Hras-1

Rnu2pa-Hras1 Hras-I-Igflr

&flr

Igfl r-Fes Fes-Tyr Tyr-Emu-23 Emu-23-D7Wasl2 D’/WaslB-Calc Calc-Pth Pth-Hbb Hbb-H-19 H-19-Th Th-Fis-1

Mag

Fes Tyr Emu-23 D7Wasl2 Calc Pth Hbb H-19 Th Fis-1

a Human locus symbols and chromosomal * Human homolog has not been determined. ’ Provisional symbol designation.

-

7/334 o/334 l/114 o/114 12/114 o/114 o/114

0.9

0.02

0.0

0.00

10.5 0.0

4.42 0.00

TGFBl

lSq13.1

Dl9FllSl MAG GPI

lSq13.1 lSq13.1 lSq13.1

N.D.” N.D.”

3.07

LHB

0.0

0.00

3.07

N.D.b

o/114 o/114

0.0 0.0

0.00 0.00

3.07

N.D.6

3.07

KLKl

lSq13.3-qter

o/114

0.0

0.00

3.07

RRAS

lSq13.3-qter

l/114 11/114

0.9

9.6

0.02 3.81

4.62 14.48

SNRP70 MYODl

lSq13.3-qter llp15.4

2/114 o/114

1.8 0.0

0.21 0.00

5.96

N.D.b

4.62

HRASl

llp15.5

71114 5/114 l/114 l/114 a/114 o/114 l/114 171114 l/114 l/114

6.1 4.4 0.9 0.9 7.0 0.0 0.9 14.9 0.9 0.9

1.85 1.15 0.02 0.02 2.39 0.00 0.02 7.18 0.02 0.02

10.76 9.03 4.62 4.62 11.85 3.07 4.62 19.82 4.62 4.62

IGFl R FES TYR

15q25-qter 15q25-qter llq14-q21

lSq13.3

N.D.b N.D.b

CALC PTH HBB

llp15.4 llpter-p15.5 llp15.5

N.D.”

TH

llp15.5

N.D.6 assignments

(14,36,48).

gld chromosomes, which may cause a suppression of recombination. The murine t complex is the only reported example of inversions that resulted in a 50- to loo-fold suppression of recombinations (Hammer et al., 1989). The interspecific backcross analysis reported here identified seven clusters of closely linked loci (Le., there were no recombinants in either the 334

or 114 progeny analyzed).

These clusters were (1)

Apoe, Atpa-2, Ckmm, D19S19h, and Ercc-2; (2) Cyp2a, Cyp2b, Rip, and Tgfb; (3) Mag and Gpi; (4) D7Rp2 and Cebp; (5) Egfbp, Kal, Lhb, Ngfg, Rras, and Rnulp70; (6) Hras-1 and Igflr; and (7) Calc and Pth. We previously reported that the interval between and Cyp2b/C YP2A is compressed from

Pkcg/PRKCG

532

SAUNDERS

33.1 CM in humans to 3.3 CM in the interspecific backcross mice. In addition, we did not observe any recombinants in the 334 mice studied between the mouse homologs of APOE, ATPlA3, CKMM, D19S19, and ERCC2 (Saunders and Seldin, 1990). In humans, these loci have been individually ordered with respect to DM and span a genetic distance of approximately 5 CM (Keats et al., 1989; B. Keats, personal communication). Recombination may also be suppressed between Calc and Pth. Two human genetic linkage analyses separate CALC and PTH by approximately 8 CM; a third report found no recombinants in data on 130 phase-known meioses (reviewed by van Heyningen and Porteus, 1986). A recent study of mitotic deletions of human chromosome llp15.5 indicates that the CALC locus is distal to the PTH locus (Henry et al., 1989). Pulsed-field gel electrophoretic studies have placed the human homologs of Cyp2a and Cyp2b on the same 350-kb fragment (Miles et aZ., 1989), suggesting that the lack of recombination between these genes in backcross mice is due to close physical proximity. TGFBl, the human homolog of Tgfb, has been colocalized with CYP2A and CYP2B by somatic cell hybrid panels (&honk et al., 1989), and crossovers between these genes in humans have not been reported (LeBeau et al., 1989). Egfbp, Kal, and Ngfg are members of the highly homologous kallikrein multigene family that contains more than 20 genes. These genes are closely linked, and the family appears to represent a single genetic locus on mouse chromosome 7 (Evans et al., 1987). In situ hybridization suggests that all human kallikrein genes are organized in a single locus on human chromosome 19 (Evans et al., 1988). Both RRAS and LHB colocalize with KLKl, the human homolog of Kal, to human chromosome 19q13.3-qter by somatic cell hybrid panels (Schonk et al., 1989).

Comparative

Mapping

of Human

Disease Loci

The mapping of homologous genes in mice and humans is useful for predicting the location of disease loci in the mouse and for identifying mouse models of human disease. In the mouse, the use of interspecific crosses circumvents the difficulty in detecting polymorphisms and the requirement in human genetics of well-characterized, extended families for accurate gene localization. In addition, a large number of genes can be evaluated at the same time. The low recombination frequencies observed in this study limit the usefulness of comparative mapping in the linear ordering of genes closely linked to DM. However, the use of interspecific backcross mouse genetics can serve as a rapid means of establishing whether new human chromosome 19-specific probes

AND

SELDIN

and candidate disease genes map to the region of interest. This is of particular value when human polymorphisms for a new probe cannot be detected or are uninformative. For example OTF2, HKRl, ERCC2, ICAMl, MAG, PVS, and XRCCl have been regionally localized to human chromosome 19 using somatic cell hybrid panels. These genes have not been accurately localized by genetic linkage analysis because informative polymorphisms have not been detected (LeBeau et al., 1989). Polymorphisms for D19S19 subclones are minimally informative (Bartlett et aZ., 1987; M. Pericak-Vance, personal communication); consequently, D19S19 has been mapped to 19cenq13.2 by only limited genetic linkage analyses. Unique M. spretus variants were easily detected for all of these probes in our panel of interspecific backcross mice, and the linkage relationship of the homologous mouse loci to the putative DM gene region was quickly established. As new human chromosome 19-specific probes and candidate genes for DM become available, their segregation relative to loci within the syntenic region of mouse chromosome 7 can be rapidly screened prior to initiating more labor intensive human gene linkage analyses. The current study has examined the linkage relationship of several loci associated with human disorders. In the syntenic region of human chromosome 19, GPI is associated with hemolytic anemia and hydrops foetalis; APOE is associated with familial hyperlipoproteinemia III and dysbetalipoproteinemia; and LHB is associated with fertile eunuchoidism and isolated LH deficiency (Harper et al., 1989). On the basis of the human chromosome 19q13.1-q13.4 homology, the prediction that the human disease loci for myotonic dystrophy (DM), a form of xeroderma pigmentosum (ERCCI), and hyperlipoproteinemia Ib (APOC2) would map to the proximal portion of mouse chromosome 7 can be postulated. A number of clinical disorders have been associated with HBB, which is found on human chromosome 11. These disorders include P-thalassemia, sickle cell anemia, @-methemoglobinemia, and Heinz body anemia (Harper et aZ., 1989). PTH, associated with familial hypoparathyroidism, and TYR, linked to tyrosinasenegative albinism, are also found on human chromosome 11 (Harper et aZ., 1989). Since the comparative gene order between mouse chromosome 7 and human chromosome 11 has not been grossly conserved, the localization of the numerous other diseases associated with human chromosome llp15.4-~15.5 and llq14q21 cannot be accurately predicted at the present time. Future studies delineating the borders of what may be relatively small conserved linkage groups should facilitate the process of predicting where homologous human chromosome 11 disease loci are located in the mouse.

LINKAGE

MAP

OF

ACKNOWLEDGMENTS We thank the following people for donating probes: A. Lusis (Apoe), R. Richards (APS and Kal), R. Levenson (Atpa-2), T. McKeithan (BCL3), J. Jacobs (C&c), K. Xanthopoulos (Cebp), F. Gonzalez (CYP2A), C. Kasper (Cyp2b), G. Watson (D7Rp2), D. Adler and C. Disteche (D7Wus12), A. Roses and J. Gilbert (DZ9SZ9), K. Johnson (D19S51), M. Blaber (Egfbp), E. Rinchik (Emu-23), M. van Duin (ERCCI), J. Silver (Fi.s-I), M. Gurney and T. Siddique (GPO, B. Vogelstein (HKRZ and HKRP), S. Tilghman (H-191, A. Bryan and P. Kuhlman (Icon-Z), J. Roder (Mug), H. Weintraub (MYODZ), L. Staudt (OTF2), K. Gross (Ngfg), G. Heinrich (Pth), V. Racaniello (PVS), M. Negishi (Rip), A. Singh (Rrus), J. Keene (RnuZp70 and Rnu2pa), R. Stearman (&o-3), B. Kwon (Tyr), R. Derynck (Tgfb), and L. Thompson, M. Siciliano, and L. Bachinski (XRCCI). The funding for this work was provided by the Muscular Dystrophy Association of America and NIH Grants NS19999 and HGOOlOl. Note added in proof. The provisional mouse symbols for the loci listed as Rnulp70 and Rnu2pu have now been changed to Snrp70 and Snrp2u, to correspond with the human nomenclature.

MOUSE

CHROMOSOME 11.

CECI, J. D., SIRACUSA, L. D., JENKINS, N. A., AND COPELAND, N. G. (1989). A molecular genetic linkage map of mouse chromosome 4 including the localization of several proto-oncogenes. Genomics 6: 699-709.

12.

CECI, J. D., JUSTICE, M. J., LOCK, L. F., JENKINS, N. A., AND COPELAND, N. G. (1990). An interspecific backcross linkage map of mouse chromosome 8. Genomics 6: 72-79.

13.

COUSSENS, L., PARKER, P. J., RHEE, L., YANG-FEN, T. L., CHEN, E., WATERFIELD, M. D., FRANCKE, U., AND ULLRICH, A. (1986). Multiple, distinct forms of bovine and human protein kinase C suggest diversity in cellular signaling pathways. Science 233: 859-866.

14.

Cox, D. W., AND DONLON, T. A. (1989). Report of the committee on the genetic constitution of chromosomes 14 and 15: Human Gene Mapping 10. Cytogenet. Cell Genet. 51: 280-298.

15.

DANIEL, W. W. (1987). “Biostatistics: A Foundation for Analysis in the Health Sciences,” 4th ed., pp. 531-538, Wiley, New York.

16.

DAVIS, R. L., WEINTRAUB, H., AND LASSER, A. B. (1987). pression of a single transfected cDNA converts fibroblasts myoblasts. Cell 61: 987-1000.

17.

DAVISSON, M. T., RODERICK, T. H., HILLYARD, A. L., AND DOOLI’ITLE, D. P. (1989). The locus map of the mouse. Mouse News Z&t. 84: 15-23.

18.

DERYNCK, R., JARRET, J. A., CHEN, V. (1986). The murine transforming sor. J. Biol. Chem. 261: 4377-4379.

19.

DISTECHE, C. M., AND ADLER, D. (1984). Localization of cloned mouse chromosome 7-specific DNA to lethal albino deletions. Somatic Cell Mol. Genet. 10: 211-215.

20.

DULL, T. J., GRAY, A., HAYFLICK, J. S., AND ULLRICH, A. (1984). Insulin-like growth factor II precursor gene organization in relation to insulin gene family. Nature Ukndon) 310: 777-781.

21.

ELLIS, R. W., DEFEO, D., SHIH, T. Y., GONDA, M. A., YOUNG, H. A., TSUCHIDA, N., Lowy, D. R., AND SCOLNICK, E. M. (1981). The p21 src genes of Harvey and Kirsten sarcoma viruses originate from divergent members of a family of normal vertebrate genes. Nature (London) 292: 506-511.

22.

EVANS, B. A., DRINKWATER, C. C., AND RICHARDS, R. I. (1987). Mouse glandular kallikrein genes: Structure and partial sequence analysis of the kallikrein gene locus. J. Biol. Chem. 262: 8027-8034.

23.

EVANS, B. A., YUN, 2. X., CLOSE, J. A., TREGEAR, G. W., KITAMURA, N., NAKANISHI, S., CALLEN, D. F., BAKER, E., HYLAND, V. J., SUTHERLAND, G. R., AND RICHARDS, I. R. (1988). Structure and chromosomal localization of the human renal kallikrein gene. Biochemistry 27: 3124-3129. FERRIS, S. D., SAGE, R. D., AND WILSON, A. C. (1982). Evidence from mtDNA sequences that common laboratory strains of inbred mice are descended from a single female. Nature (London) 295: 163-165. FRESCO, L. D., KURILLA, M. G., AND KEENE, J. D. (1987). Rapid inhibition of processing and assembly of small nuclear ribonucleoproteins after infection with vesicular stomatitis virus. Mol. Cell. Biol. 7: 1148-1155.

REFERENCES 1.

AVNER, (1988). crosses.

2.

BARTLE?T, R. J., PERICAK-VANCE, M. A., YAMAOKA, L., GILBERT, J., HERBSTREITH, M., HUNG, W.-Y., LEE, J. E., MoHANDAS, T., BRUNS, G., LABERGE, C., THIBAULT, M.-C., Ross, D., AND ROSES, A. D. (1987). A new probe for the diagnosis of myotonic muscular dystrophy. Science 236: 16481650. BARTON, D. E., ARQUINT, M., RODER, J., DUNN, R., AND FRANCKE, U. (1987). The myelin-associated glycoprotein gene: Mapping to human chromosome 19 and mouse chromosome 7 and expression in quivering mice. Genomics 1: 107112. BELL, G. I., HORITA, S., AND KARAM, J. H. (1984). A polymorphic locus near the human insulin gene is associated with insulin-dependent diabetes mellitus. Diabetes 33: 176-183. BERGER, F. G., GROSS, K. W., AND WATSON, G. (1981). Isolation and characterization of a DNA sequence complementary to an androgen-inducible messenger RNA from mouse kidney. J. Biol. Chem. 256: 7006-7013.

3.

4.

5.

P., AMAR, L., DANDOLO, Genetic analysis of the Trends Genet. 4: 18-23.

L., AND GUENET, J. L. mouse using interspecific

6.

BISHOP, D. T. (1985). The information content of phaseknown matings for ordering genetic loci. Genet. Epidemiol. 2: 349-361.

24.

7.

BLABER, M., ISACKSON, P. J., AND BF~ADSHAW, R. A. (1987). A complete cDNA sequence for the major epidermal growth factor binding protein in the male mouse submandibular gland. Biochemistry 26: 6742-6749.

25.

8.

BLANK, R. D., CAMPBELL, G. R., CALABRO, A., AND D’EUSTACHIO, P. (1988). A linkage map of mouse chromosome 12: Localization of Zgh and effects of sex and interference on recombination. Genetics 120: 1073-1083.

26.

9.

BONHOMME, F., CATALAN, J., BRIITON-DAVIDIAN, MAN, V., MORIWAKI, K., NEVO, E., AND THALER, Biochemical diversity and evolution in the genus them. Genet. 22: 275-303.

10.

J., CHAPL. (1984). Mus. Bio-

BUCKLE, V. J., EDWARDS, J. H., EVANS, E. P., JONASSON, J. A., LYON, M. F., PETER, J., AND SEARLE, A. G. (1985). Comparative maps of human and mouse X chromosomes: Human Gene Mapping 8. Cytogenet. Cell Genet. 40: 593-594.

533

7

27.

28.

EXto

E. Y., AND GOEDDEL, D. growth factor-p precur-

GREEN, E. L. (1981). Linkage, recombination and mapping. In “Genetics and Probability in Animal Breeding Experiments” (E. Green, Ed.), pp. 77-113, Macmillan, New York. GREVE, J. M., DAVIS, G., MEYER, A. M., FORTE, C. P., YOST, S. C., MARLOR, C. W., KAMARK, M. E., AND MCCLELLAND, A. (1989). The major human rhinovirus receptor is ICAM-1. Cell 66: 839-847. HAMMER, M. F., SCHIMENTI, J., AND SILVER, L. M. (1989). Evolution of mouse chromosome 17 and the origin of inver-

534

SAUNDERS sions associated 86: 3261-3265.

29.

with

t haplotypes.

Proc. Natl.

Acad.

HARPER, P. S., FREZAL, J., FERGUSON-SMITH, M. A., AND SCHINZEL, A. (1989). Report of the committee on clinical disorders and chromosomal deletion syndromes: Human Gene Mapping 10. Cytogenet. Cell Genet. 51: 563-611.

locus.

Proc.

Natl.

45.

LALLEY, P. A., SAKAGUCHI, A. Y., EDDY, R. L., HONEY, N. H., BELL, G. I., SHEN, L.-P., RU?TER, W. J., JACOBS, J. W., HEINRICH, G., CHIN, W. W., AND NAYLOR, S. L. (1987). Mapping of polypeptide hormone genes in the mouse: Somatostatin, glucagon, calcitonin, and parathyroid hormone. Cytogenet. Cell Genet. 44: 92-97.

46.

LALLEY, P. A., DAVISSON, M. T., GRAVES, J. A. M., O’BRIEN, S. J., WOMACK, J. E., RODEFUCK, T. H., CREAU-GOLDBERG, N., HILLYARD, A. L., DOOLI~LE, D. P., AND ROGERS, J. A. (1989). Report of the committee on comparative mapping: Human Gene Mapping 10. Cytogenet. Cell Genet. 61: 503-532.

47.

LAWN, R. M., FRITSCH, E. F., PARKER, R. C., BLAKE, G., AND MANIATIS, T. (1978). The isolation and characterization of 6and @-globin genes from a cloned library of human DNA. Cell 15: 1157-1174.

48.

HUMPHERIES, S. E., BERG, K., GILL, L., CUMMING, A.M., ROBERTSON, F. W., STALENHOEF, A. F. H., WILLIAMSON, R., AND BORREXON, A. L. (1984). The locus for apolipoprotein C-II is closely linked to the gene for apolipoprotein E on chromosome 19. Clin. Genet. 26: 389-396.

LE BEAU, M. M., RYAN, D., AND PEFUCAK-VANCE, M. A. (1989). Report of the committee on the genetic constitution of chromosomes 18 and 19: Human Gene Mapping 10. Cytogenet. Cell Genet. 61: 338-357.

49.

JACOBS, J. W., GOODMAN, R. H., CHIN, W. W., DEE, P. C., HABENER, J. F., BELL, N. H., AND Pms, J. T., JR. (1981). Calcitonin messenger RNA encodes multiple polypeptides in a single precursor. Science 213: 457-459.

LOWE, D. G., CAPON, D. J., DELWART, E., SAKAGUCHI, A. Y., NAYLOR, S. L., AND GOEDDEL, D. V. (1987). Structure of the human and murine r-rus genes, novel genes closely related to ras proto-oncogenes. Cell 48: 137-146.

50.

JANDEL, A. S., ICKING, D., KABISCH, R., OLEK, K., DRIESEL, A., AND GRZESCHIK, K. H. (1985). Human cDNA probes detecting RFLPs on human chromosomes. Cytogenet. Cell Genet. 40: 660.

Lusrs, A. J., TAYLOR, B. A., QUON, D., ZOLLMAN, S., AND LEBOUEF, R. C. (1987). Genetic factors controlling structure and expression of apolipoproteins B and E in mice. J. Biol. Chem. 262: 7594-7604.

51.

LYON, M. F. (1989). Lett. 84: 24-25.

52.

MCKEITHAN, T. W., ROWLEY, J. D., SHOWS, T. B., AND DIAZ, M. 0. (1987). Cloning of the chromosome translocation breakpoint junction of the t(14;19) in chronic lymphocytic leukemia. Proc. Natl. Acad. Sci. USA 84: 9257-9260.

53.

MILES, J. S., BICKMORE, W., BROOK, J. D., MCLAUREN, A. W., MEEHAN, R., AND WOLF, C. R. (1989). Close linkage of the human cytochrome P450IIA and P450IIB gene subfamilies: Implications for the assignment of substrate specificity. Nucleic Acids Res. 17: 2907-2917.

54.

MOSELEY, W. S., AND SELDIN, M. F. (1989). Definition of mouse chromosome 1 and 3 gene linkage groups that are conserved on human chromosome 1: Evidence that a conserved linkage group spans the centromere of human chromosome 1. Genomics 6: 899-905.

55.

NADEAU, J. H. (1989). Maps of linkage and synteny homologies between mouse and man. Trends Genet. 5: 82-86. NIGRO, J. M., SCHWEINFEST, C. W., RAJKOVIC, A., PAVLOVIC, N., JAMAL, S., DOTTIN, R. P., HART, J. T., KAMARCK, M. E., RAE, P. M., CARTY, M. D., AND MARTIN-DELEON, P. (1987). cDNA cloning and mapping of the human creatine kinase M gene to 19q13. Amer. J. Hum. Genet. 40: 115-125. NIMMO, E., WILLIAMSON, R., AND JOHNSON, K. (1989). Localization of c-MEL gene to 19(cen-p13.2). Cytogenet. Cell Genet. 61: 1053.

HENRY, I., GRANDJOUAN, S., BARICHARD, F., HUERRE-JEANPIERRE, C., AND JUNIEN, C. (1989). Mitotic deletions of 11~15.5 in two tumors indicate that the CALCA locus is distal to the PTH locus. Cytogenet. Cell Genet. 50: 155-157.

32.

HOWLES, P. N., DICKINSON, D. P., DICAPRIO, L. L., WOODWORTH-GUTAI, M., AND GROSS, K. W. (1984). Use of a cDNA recombinant for the gamma subunit of murine nerve growth factor to localize members of this multigene family near the TAM-l locus on chromosome 7. Nucleic Acids Res. 12: 27912805.

36.

JUNIEN, C., AND MCBRIDE, 0. W. (1989). Report of the committee on the genetic constitution of chromosome 11: Human Gene Mapping 10. Cytogenet. Cell Genet. 51: 226-258.

37.

KEATS, B., OTT, J., AND CONNEALLY, M. (1989). Report of the committee on linkage and gene order: Human Gene Mapping 10. Cytogenet. Cell Genet. 51: 459-502.

38.

KENT, R. B., FALLOWS, D. A., GEISSLER, E., GLASER, T., EMANUEL, J. R., LALLEY, P. A., LEVENSON, R., AND HOUSMAN, D. E. (1987). Genes encoding a and @ subunits of Na,K-ATPase are located on three different chromosomes in the mouse. Proc. Natl. Acad. Sci. USA 84: 5369-5373.

39.

c-albino

LALLEY, P. A., AND CHIRGWIN, mouse insulin genes. Cytogenet.

31.

35.

tyrosinase that maps at the mouse Acad. Sci. USA 84: 7473-7477. 44.

HEINRICH, G., KRONENBERG, H. M., POTTS, J. T., JR., AND HAEIENER, J. F. (1984). Gene encoding parathyroid hormone: Nucleotide sequence of the rat gene and deduced amino acid sequence of rat preproparathyroid hormone. J. Biol. Chem. 259: 3320-3329.

34.

SELDIN

Sci. USA

30.

33.

AND

KINGSLEY, D. M., JENKINS, N. A., AND COPELAND, N. G. (1989). A molecular genetic linkage map of mouse chromosome 9 with new localizations for the G&a, T3q, E&-l, and Ldlr loci. Genetics 123: 165-172.

40.

KNOTT, T. J., EDDY, R. L., ROBERTSON, M. E., PRIESTLEY, L. M., Scorr, J., AND SHOWS, T. B. (1984). Chromosomal localization of the human apoprotein CI gene and of a polymorphic apoprotein AI1 gene. Biochem. Biophys. Res. Commun. 126: 299-306.

56.

41.

Ko, H., FAST, P., MCBRIDE, W., AND STAUDT, L. M. (1988). A human protein specific for the immunoglobulin octamer DNA motif contains a functional homeobox domain. Cell 66: 135-144.

57.

42.

KORNELUK, R. G., MACLEOD, H. L., MCKEITHAN, T. W., BROOKS, J. D., AND MACKENZIE, A. E. (1989). A chromosome 19 clone from a translocation breakpoint shows close linkage and linkage disequilibrium with myotonic dystrophy. Genomics 4: 146-151.

43.

KWON, B. S., HAQ, A. K., POMERANTZ, S. H., AND HALABAN, R. (1987). Isolation and sequence of a cDNA clone for human

58.

59.

Mouse

J. M. (1984). Mapping Cell Genet. 37: 515.

chromosome

atlas.

Mouse

of

News

NOSHIRO, M., LAKSO, M., KAWAJIRI, K., AND NEGISHI, M. (1988). Rip locus: Regulation of female specific isozyme (I-P450,,) of testosterone 16cY-hydroxylase in mouse liver, chromosome localization, and cloning of P-450 cDNA. Biochemistry 27: 6434-6443. O’MALLEY, K. L., AND ROT-IN, P. (1988). Human tyrosine hydroxylase and insulin genes are contiguous on chromosome 11. Nucleic Acids Res. 16: 4437-4446.

LINKAGE

MAP

OF

60.

PACHNIS, V., BELAYEW, A., AND TILGHMAN, S. M. (1984). Locus unlinked to cY-fetoprotein under the control of the murine Raf and Rif genes. Proc. Natl. Acad. Sci. USA 81: 5523-5527.

61.

PADUA, R. A., BARRAS, N., AND Cum, transforming gene in a human malignant Nature (London) 311: 671-673.

CHROMOSOME 76.

G. A. (1984). A novel melanoma cell line.

62.

QUERY, C. C., Am KEENE, J. D. (1987). A human autoimmune protein associated with Ul RNA contains a region of homology that is cross-reactive with retroviral~30~ antigen. Cell 51: 211-220.

63.

RICHARDS, R. I., CATANZORO, D. F., MASON, A. J., MORRIS, B. J., BAXTER, J. D., AND SHINE, J. (1982). Mouse glandular kallikrein genes. J. Biol. Chem. 267: 2758-2761.

64.

RINCHIK, E. M., MACHANOFF, R., CUMMINGS, C. C., AND JOHNSON, D. K. (1989). Molecular cloning and mapping of the ecotropic leukemia provirus Emv-23 provides molecular access to the albino deletion complex in mouse chromosome 7. Genomics 4: 251-258.

65.

MOUSE

7

535

SILVER, J., AND BUCKLER, C. E. (1986). A preferred region for the integration of Friend murine leukemia virus in hematopoietic neoplasms is closely linked to the Znt-2 oncogene. J. Virol. 60: 1156-1158. SIMMONS, D. L., LALLEY, P. A., AND KASPER, C. B. (1965). Chromosomal assignments of genes coding for components of the mixed-function oxidase system in mice: Genetic localization of the cytochrome P-45OPCN and P-450PB gene families and the NADPH-cytochrome P-450 oxidoreductase and epoxide hydratase genes. J. Biol. Chem. 260: 515-521.

78.

SIRACUSA, L. D., BUCHBERG, A. M., COPELAND, N. G., AND JENKINS, N. A. (1989). Recombinant inbred strain and interspecific backcross analyses of molecular markers flanking the agouti color locus. Genetics 122: 669-679.

79.

SODROSKI, J. G., GOH, W. C., AND HASELTINE, W. A. (1984). Transforming potential of a human protooncogene (c-fps/fes) locus. Proc. Natl. Acad. Sci. USA 81: 3039-3043.

80.

STEARMAN, R. S., LOWELL, C. A., PELTZMAN, C. G., AND MORROW, J. F. (1986). The sequence and structure of a new serum amyloid A gene. Nucleic Acids Res. 14: 797-809.

RUPPERT, J. M., KINZLER, K. W., WONG, A. J., BIGNER, S. H., Kno, F., LAW, M. L., SEUANEZ, H. N., O’BRIEN, S. J., AND VOGELSTEIN, B. (1988). The GLZ-Kruppel family of human genes. Mol. Cell. Biol. 8: 3104-3113. RYDER-COOK, A. S., SICINSKI, P., THOMAS, K., DAVIES, K. E., WORTON, R. G., BARNARD, E. A., DARLISON, M. G., AND BARNARD, P. J. (1988). Localization of the mdx mutation with the mouse dystrophin gene. EMBO J. 7: 3017-3021.

81.

SUTHERLAND, G. R., BAKER, E., HYIAND, V. J., CALLEN, D. F., CLOSE, J. A., TREGEAR, G. W., EVANS, B. A., AND RICHARDS, E. I. (1988). Human prostate-specific antigen (APS) is a member of the glandular kallikrein gene family at 19ql3. Cytogenet. Cell Genet. 48: 205-207.

82.

67.

SAUNDERS, A. M., AND S&N, M. F. (1990). The syntenic relationship of proximal mouse chromosome 7 and the myotonic dystrophy gene region on human chromosome 19q. Genomics 6: 324-332.

TALMADGE, K., BOORSTEIN, W. R., AM) FIDDES, J. C. (1983). The human genome contains seven genes for the &subunit of chorionic gonadotropin but only one gene for the p-subunit of luteinizing hormone. DNA 2: 281-289.

83.

68.

SCHONK, D., COERWINKEL-DRIESSEN, M., VAN DALEN, I., OERLEMANS, F., Smm, B., SCHEPENS, J., HULSEBOS, T., COCKBURN, D., BOYD, Y., DAVIS, M., RUG, W., SHAW, D., ROSES, A., ROPERS, H., AND WIEFUNGA, B. (1989). Definition of subchromosomal intervals around the myotonic dystrophy gene region at 19q. Genomics 4: 384-396.

ULLRICH, A., GRAY, A., TM, A. W., YANG-FEN, T., Tsv~oKAWA, M., COLLINS, C., HENZEL, W., LE BON, T., KATHUIUA, S., CHEN, E., JACOBS, S., FRANKS, U., RAMACHANDRAN, J., AND FUJITA-YAMAGUCHI, Y. (1986). Insulin-like growth factor I receptor’s primary structure: Comparison with insulin receptor suggests structural determinants that define functional specificity. EMBO J. 5: 2503-2512.

69.

SEARLE, A. G., PETERS, J., LYON, M. F., EVANS, E. P., EDWARDS, J. H., AND BUCKLE, V. J. (1987). Chromosome maps of man and mouse, III. Genomics 1: 3-18. SELDIN, M. F., MORSE, H. C., REEVES, J. P., SCRIBNER, C. L., LEBOELJF, R. C., AND STEINBERG, A. D. (1988). Genetic analysis of “autoimmune” gld mice. I. Identification of a restriction fragment length polymorphism closely linked to the gld mutation within a conserved linkage group. J. Exp. Med. 167: 688-693. SELDIN, M. F., HOWARD, T. A., AND D’EUSTACHIO, P. (1989). Comparison of linkage maps of mouse chromosome 12 derived for laboratory strain intraspecific and Mus spretus interspecific backcrosses. Genomics 5: 24-28.

84.

VAN DUIN, M., DE Wm, J., ODIJK, H., WEBTER~ELD, A., YASUI, A., KOKEN, M., HOEIJMAKERS, J., AND BOOTSMA, D. (1986). Molecular characterization of the human excision repair gene ERCC-I: cDNA cloning and amino acid homology with the yeast DNA repair gene RADlO. Cell 44: 913-923.

85.

VAN HEYNINGEN, V., AND PORTEUS, D. J. (1986). Mapping a chromosome to find a gene. Trends Genet. 2: 4-5. WEBER, C. A., SALAZAR, E. P., STEWART, S. A., AND THOMPSON, L. H. (1988). Molecular cloning and biological characterization of a human gene, ERCCZ, that corrects the nucleotide excision repair defect in CHO UV5 cells. Mol. Cell Biol. 8: 1137-1146.

66.

70.

71.

72.

SMW, D. J., MERIDITH, A. L., SARFARZI, M., HARLEY, H. G., HUSON, S. M., BROOK, J. D., BUFTON, L., Lrrr, M., MOHANDAS, T., AND HARPER, P. S. (1986). Regional localization and linkage relationships of seven RFLPs and myotonic dystrophy on chromosome 19. Hum. Genet. 74: 262-266.

73.

SMW, D. J., AND EIBERG, H. (1987). Report of the committee for chromosomes 17, 18, and 19: Human Gene Mapping 9. Cytogenet. Cell Genet. 46: 242-256.

74.

SICILIANO, M. J., CARRANO, A. V., AND THOMPSON, L. H. (1986). Assignment of a human DNA-repair gene associated with sister-chromatid exchange to chromosome 19. Mut. Res. 174: 303-308. SIDDIQUE, T., MCKINNEY, R., HUNG, W., Bmm, R., BRUNS, G., MOHANDAS, T. K., ROPERS, H., WILFERT, C., AM) ROSES, A. D. (1988). The poliovirus sensitivity (PVS) gene is on chromosome 19ql2-q13.2. Genomics 3: 156-160.

75.

86.

87.

XANTHOPOULOS, K. G., MIRKOVITCH, J., FRIEDMAN, J. M., AND DARNELL, J. E., JR. (1989). The mouse Cebp gene encoding a DNA-binding protein is polymorphic and is located on chromosome 7. Cytogenet. Cell Genet. 50: 174-175.

88.

YAMANO, S., NAGATA, K., YAMAZOE, Y., KATO, R., GELBOIN, H., AND GONZALEZ, J. (1989). cDNA and deduced amino acid sequences of human P450 IIA3 (CYP2A). Nucleic Acids Res. 17: 4888. YANG-FENG, T. L., DEGENNARO, L. J., AND FRANKE, U. (1986). Genes for synapsin I, a neuronal phosphoprotein, map to conserved regions of human and murine X chromosomes. Proc. Natl. Acad. Sci. USA 83: 8679-8683.

89.

90.

ZIMMERMAN, W., ORTLIEB, B., FRIEDRICH, R., AND VON KLEIST, S. (1987). Isolation and characterization of cDNA clones encoding the human carcinoembryonic antigen reveal a highly conserved repeating structure. Proc. Natl. Acad. Sci. USA 84: 2960-2964.