Ann. Hum. Genet., Lond. (1977), 41, 61

61

Printed i n Great Britain

The linkage relationships of the haemoglobin beta, delta and alpha loci with 34 genetic marker systems BY L. R. WEITKAMP,* G. STAiUTOYANNOPOULOS,t P. T. ROWLEY2 AND R. L. KIRK$ Two haemoglobins, Hb A and HbA,, are easily detectable on electrophoresis of haemolysates from adults. Haemoglobin A is a tetramer composed of two a and two B chains; haemoglobin A, of two a and two 6 chains. The major foetal haemoglobin, Hb F, consists of two a and two y chains. Recent studies have indicated that multiple loci are involved in the production of y and possibly a chains, but the /3 and S chains may be specified by only one locus each (for review see Stamatoyannopoulos & Nute, 1974; Schwartz, 1976). The haemoglobin a and p loci are not closely linked (Table 3 for review). Indeed, from in situ mRNA hybridization (Price, Conover & Hirschhorn, 1972) and segregation in somatic cell hybrids (Deisseroth, Velez & Nienhuis, 1976) these loci are probably on different chromosomes. Based on the structures of the 6/?hybrids in Lepore haemoglobins (Baglioni, 1962) and a lack of recombination between the @ and 6 loci (Mishu & Nance, 1969), the 6 and /3 loci are closely linked. The structure of haemoglobin Kenya, a ?@ fusion chain, indicates that the 9 and /3 loci are closely linked (Huisman, Miller & Schroeder, 1972; Kendall et al. 1973). Although haemoglobin is the most thoroughly investigated component of blood, the chromosomal location and linkage relationships of the haemoglobin loci remain unclear. Price et al. (1972) suggested that one group of haemoglobin loci may be on chromosome no. 2 and the other on a B-group chromosome. This assignment is apparently not confirmed using cDNA-DNA hybridization in somatic cell hybrid clone lines (Anderson et al. 1976). Family studies of the linkage relationships of the haemoglobin ,8 and 6 loci with up to 25 marker systems (Morganti, Beolchini & Gualandri, 1969; Nance et al. 1970; Edwards, 1974; Falk et al. 1975) have not provided significant evidence favouring linkage with any of the loci tested. A study of the Hopkins-2 a chain variant also failed to yield significant evidence of linkage with any of seven marker systems (Bradley, Boyer & Allen, 1961). Here we report a study of the linkage relationships of the haemoglobin @-6-yregion in 151 families with 34 informative marker systems, including 13 loci for which data have not been previously reported. The results of analysis of the linkage relationships of the haemoglobin a locus in 21 families with 19 informative marker systems are also presented. MATERIAL AND METHODS

The families analysed for the linkage relationships of the haemoglobin alpha and beta loci nclude 96 families resident in Greece, 3 from New Guinea, one from Haiti and 70 from Rochester, New York. The first three groups were ascertained during the course of studies for other purposes,

* Department of Psychiatry, Division of Genetics, and Department of Pediatrics,University of Rochester School of Medicine and Dentistry, Rochester, New York. t Division of Medical Genetics, Department of Medicine, University of Washington School of Medicine, Seattle, Washington. $ Departments of Medicine and Pediatrics, Division of Genetics and Department of Microbiology, University of Rochester School of Medicine and Dentistry, Rochester, New York. $ John Curtin School of Medical Research, Australian National University, Canberra, Australia.

62

L. R.WEITKAMP AND

OTHERS

but the Rochester families were selected primarily for segregation a t the haemoglobin beta locus and include a number of third-generation phase-known offspring. Individuals informative for the linkage relationships of the P-S-y region include 79 persons heterozygous for P-thalassaemia; six of these were from an Italian family in Rochester, one from a Rochester black family, and the rest were residents of Greece. One additional Greek family was segregating for pS thalassaemia. The diagnosis of p or pS thalassaemia in Greek families was based on quantitations of Hb A, and Hb F, evaluations of red cell morphology, and red cell indices (MCH and MCV), using described methodology and diagnostic criteria (Malamos, Fessas & Stamatoyannopoulous, 1962). I n the Rochester families thalassaemia phenotypes were determined by A, quantitation using isoelectric focusing (Rowley et al. 1972), osmotic fragility and peripheral blood smear morphology. I n one Greek family the genotype of the untested heterozygous individual was inferred from the phenotypes of his wife and six offspring. I n all other cases both the informative heterozygote and his (her) spouse were directly typed. Seventy-two individuals were segregating for the haemoglobin p-6-y region electrophoretic variants S, C, B, or hereditary persistence of foetal haemoglobin (HPFH). Thesc include 7 Greek, 1 Haiti, 2 Rochester white and 62 Rochester black families. Haemoglobin electrophoretic variants with mobility indistinguishable from S, C, B, or F on alkaline starch-gel electrophoresis (Efremov et aE. 1969) were considered variants of the p-S-y region, providing that in the case of variants with mobility indistinguishable from S they were also positive in the dithionite tube test (Nalbandian et al. 1971). For 68 of the 72 heterozygous individuals the spouse was also typed. I n the remaining four cases in which the spouse of the indiviclual heterozygous for haemoglobin was not tested, the only offspring scored in the linkage calculations were those for whom no assumption about the genotype of the untested parent was necessary. For alpha haemoglobin variants the sample consists of 16 Greek families (alpha thalassaemia), three families of New Guinea indigenes from Kar Kar Island (haemoglobin J Tongariki) and three informative individuals from a three-generation American Negro family. The variant in the latter family, a non-sickling haemoglobin with electrophoretic mobility indistinguishable from H b S, was determined t o be an a chain variant by hybridization with known alpha and beta chain variants. The diagnosis of a thalassaemia was based on finding rare erythrocytes with H b H inclusions in individuals having erythrocyte morphology of heterozygous thalassaemia, low MCH values, normal or elevated serum iron measurements, and normal results of quantitations of H b A, and H b F (Malamos et a2. 1962). I n all cases both parents in each family were tested. The Greek families were typed for 8 red-cell antigenic loci, specifically ABO (A,, A,, B), R h (C, C", C, D , E, e), MN (M, N, S, s), Kell (K), P (P,) Lutheran (Lu"), Secretor (inferred from red cell Lea type), Duffy (Pya), 6 serum protein loci, transferrin (Tf), haptoglobin (Hp,), cholinesterase E, and E,, group-specific component (Gc), and Gm immunoglobulins, and 3 rcdcell enzyme loci, acid phosphatase (AcP,), 6 phosphogluconate dehydrogenase (PGD) and phosphoglucomutase (PGM,). The Rochester families were typed for 11 red-cell and salivary antigenic loci, namely ABO (A,, A,, B), Rh (C, c, D, D", E, e), MN (M, N, S, s ) , Kell (K, Kpa, Js*), P (P,),Lutheran (ha), Lewis (Lea, Leb, salivary Lea), Secretor (salivary H), Duffy (Fy", Fyb) and Kidd (Jk",Jkb), the white-cell antigenic loci, HLA-A and B, the 2 parotid fluid loci P b and Pr (Db), salivary amylase (Amy,), 8 serum protein loci, Tf, Hpa, cholinesterase El, Gc, properdin factor B (Bf), ceruloplasmin (Cp), complement C3 (C3), and alpha,-antitrypsin

Linkage relationships of haemoglobin loci with genetic marker systems

63

(Pi), 9 red cell enzyme loci, AcP,, PGD, PGM,, PGM,, galactose-1-phosphate uridyl transferase (Gt), peptidase A (pep A), adenosine deaminase (ADA), glutamic-pyruvic transaminase (GPT) and carbonic anhydrase I1 (CA 11) and urinary amylase (Amy,) and pepsinogen (Pg). References to standard procedures for these systems may be found in Shows (1975). Lod scores were calculated according to the method of Morton (1955), using his tables where appropriate. Scores for double intercross matings (type z4 and z5) were obtained from a set of tables made available by Dr Kirsten Fenger of the University of Copenhagen using a modification of the MOSM program (Woien, 1970). Children with genotypes incompatible with parental phenotypes were excluded from the analysis. RESULTS

Lod scores for the linkage relationships of the haemoglobin alpha locus have been listed separately for each sex, each population group, and according to whether segregation was for thalassaemia or electrophoretic variants (Appendix 1).A summary of the results showing scores separately only for the sex of the informative parent is given in Table 1. Where the phase of linkage could be determined, the number of recombinants and non-recombinants are presented separately. The linkage information from the phase-known (third-generation) data is included in the lod scores given for five values of the recombination fraction, 8, for each marker locus. Using the criterion of a lod score of + 3.0 as significant evidence favouring linkage, the data do not indicate linkage of Hb, with any of the marker loci. Linkage may be ruled out (lod score < - 2.0 both sexes combined) a t a recombination frequency, 8, of < 20 % for MN, a t 0 < 15 % for HLA, at 8 < 10 % for PGM,, ABO and Hp, and a t 8 < 5 % for AcP,. Very close linkage may be excluded for all of the remaining loci except P (each of which shows in the two generation data at least one recombinant with Hb,). Data on the linkage relationships of the haemoglobin p-6-7 region with 30 marker systems have again been given separately for each sex, each population group and according to whether segregation was for thalassemia, hereditary persistence of foetal haemoglobin (HPFH) or /3 or 6 electrophoretic variants (Appendix 2). The results for four other systems, ABO, Rh, Hp, and Gm, were not included in Appendix 2 since linkage of Hb/3 with these markers had been previously ruled out for a recombination frequency of 30 % or less (Nance et al. 1970). A summary of the lod scores for the 30 marker loci is presented separately for each sex in Table 2. There is no significant evidence favouring linkage with any of the markers tested. Linkage may be excluded (lod score < - 2.0 both sexes combined)a t 8 < 0-30for PGM, and MN, a t 8 < 0.25 for Fy, HLA, and C3, at 8 < 0.20 for PGD, AcP,, Pep A, Se, Le,P and GPT, a t 8 < 0.15 for Amy, Gt, GLO, Kell, Kidd, E, and Gc, at 8 < 0.10 for Cp and Pi and a t 8 < 0.05 for Lu, Tf, CA 11, PTC and Pr. Very close linkage is excluded for PGM,, ADA, E, and Pb, there being at east one recombinant in each instance. The results for MN include some of the families previously published (Weitkamp, Adams & Rowley, 1972),excluding those families in which the genotype of the parent scored for linkage had been inferred from his spouse and offspring.

L. R. WEITKAMP AND

64

OTHERS

Table 1. Genetic linkage relationships of the haemoglobin a locus with 19 marker systems

Chromosome I

Locus

PGD Rh PGM,

FY 2

AcP,

PhaseNo.of known hetero- offspring? zygous rparents* Nlt R 0.05

IF 2M 5F 2M 6F IM 4F

3M 6F

6

HLA GLO

9 16 Unknown

ABO HPa Lu

Gc E,

I

-088

o o

o

-3.54 -1.44 -0.19

o o

-0.65 -2.81

o

4 o

o

o

o o o

-0.72

7

0'I

-0.44 1.19 -0.65 -0.38 - 1.98 -0.88 0.05 -0.21

0'2

-0.19 0.84 -0.13 -00'02

-0.71 -0'39 0.16 0.06

0.3 -0.08

0'4 -0.02

0.47

0.14

0'01

0'01

0.14 -0.22

-0.15 0'12

0.08

0'12

-0.04 -0.04 0.04 0.03

-1-54

-0.52

-0.14

-0.02

-3.47 -2.63

-2-33

- 127

-0.64

-0.72 -0.24

-0.34

-1.56

I

-1.00

-0.70

- 0.40

-0.22

-0.10

-2'79 -2.93

-1.50

-0'43

-00'02

-0.50

-0.09

-4.33 -0.67

-2.66 -0.24

-1.16

-0.45 0.04

-O*II

-0-72

-0.44

-0.86

-0.4 -0.55

-0.19 -0.19 -0.25

-0.08 -0.08 -0.10

-0.02

-0.72

I

I

- 1.60

-0.05

0.09 0.03

a

0

gF

o

IM

o

IF

o o

o o o o o o

o

o

-1.19

-0.67

-0.25

-0.09

-0.02

o o

o o

-1.44

-0.89 0.26

-0.39

-0.15 0.17

-0.04

zF

Gm

3

o

3

IF IM

Jk

o

5M 7F

Le

P

o

1.35 -1.37

o

zF

MNSS

o o

2M 3F IF

Se(Le a)

K,Js

o o

Lod scores$ for 8 of

0'10

0'0I

0.28

0'02

-0.02 -0.02

0.05

,+M

2

2

-5.05

-3.11

-1.36

-0.53

-0.13

9F

o

-0.80

-0.36

-0.11

o

-3.60 -0.26

-2.06

2M 3F IF IF 2F IM IF

o o o

-0.07

-0.03

-0.01

-0.08

-0.01

-0.00

o o o o o

-1.19

-0.25

-0.09

-0.02

-0.02

-0.72

-0.44

-0.19 -0.07 -0.19

-0.08

-0.91

-0.17 -0.27 -0.67 -0.44 -0.42

* M = male; F = female. t NR = non-recombinant ; R

o o

o o

o o

-0.52

-0.72

0'53

0'47

0.32

0'02

-0.08

0.17

0'01 -0.02

0.05

= recombinant. $ The lod scores include the data from phase known offspring; 0 = recombination frequency.

DISCUSSION

Recombination between loci specifying haemoglobin variants was reported as early as 1967 (Schwartz et al.) and 'non-linkage' of the a and /?loci has been a standard feature of reviews on haemoglobin since 1961 (Rucknagel & Neel). We were surprised to find, however, that nowhere in the literature is there a summary of the data bearing on this question in terms of maximum-likelihood estimates of recombination frequency (lod scores). In Table 3 we present such a summary, our criterion for selection of the families being that both parents must have been typed for haemoglobin variants. Considering the total data, linkage between the Hb, and Hb, regions may be excluded for a recombination frequency of 20 yo or less.

Linkage relationships of haemoglobin loci with genetic marker systems

65

Table 2, Genetic linkage relationships of the haemoglobin /3 or 6 loci with 30 marker systems

Chromosome I

Locus

PGD PGM,

*my

PhaseNo. of known hetero- offspringt zygous parents* NR R 3M IOF zoM 25F

2M ZF

FY 2

2

AcP,

or 3

Gt

4

PGM,

6

GLO

HLA(Bf)

IZM 26 F 23M 36F IB 3F

0

0

11

11

I

8 14

10 I

2

o o I

0

II

11

I2

5

2

4

- 1.18

- 1.84

- 3.68

- 6.97 -7.49 -5.93 0.44 -3.19

3

-5.09 -0.46

-

o

0

-3.28

8F 2B 17M 34F

5

4

- 5'70

8

2M

7 15 o

-1.77 -3.31 0.33 -7.30 -10.67

11

I

3

IM

I

2

Unknown

MNSS

35M 55 F 9M 14F I B 4M 2F

5

3

25 2

21

o

0

0

3

1

0

o

0

Se§

12M

2

I

Le

I7F 13M I7F

5 o

4

4

4

0

IB 7M 14F

- 5-62 - 6.38 - 4'67

- 4'67

4

I

IB

Tf

IM

o

0

El

4 F 4M ZF

1 2

3 4

E,

ZM

o o

0 0

ZM

Go

3F 14M 32F

0 I 0

0

CP

o 3 4 1

2

0 0 0

-3.63 -1.14 -8.54 -21.48 - 1.29 -4.58 0.23 -0.42 -0.67 - 4.06 - 2'97 - 3'69 - 2'43

- 2-05 - 2.88

I

rzF

-1.12

0'12

o

0

0.23

0.18

- 3-40

I

5M 5F

- 1.18 - 7.22

0

3M

3F

0.27 -12.58 -20.73 - 1.91 -5.81 - 1.72 -17.02 -37.38 - 2.70 -8.19 0.32 - 0.91

0

IB

5

-13'42 -13'30

- 10.19 - 12.49

0

4F

c3

- 1.46 - 2'72 - 6.14 - I 1.95

- 6.01

I

ADA

Jk

- 20.82

- 1'12

o

20

P

- 1-91

- 10.05 - 17'40

IM 8M

Pep A

Lu

A

0' I

0.05

0'52

I8

K, Js, Kp

Lod scores$ for 0 of I

0'2

- 0.45 -2.51

- 4.98

>

0.3

0.4

-0.16 -0.94 -2'24

-0.22

- 5.29 - 0.66

-2.12

-0.84 -0.64

-0.38

-0.18

- 0.99

-0.52

-0.21

- 1-52

-0.57

- 2'77

- 1.02

-2.65 -0.63 0.29

-0.80

-0.13 -0.26 -0.14 0.62

-1.50

0.78 0.17 -0.69

-0'25

-0.06

-0.01

-0.00

- 1.27

-0.58

-0.16 -0.41

-0.05

0.19 -2.86

-1.02

-0.25

-2.82

-OYX

-0.45 -1.69 -0.59

-0.16 -0.77 -0.30 0.06 -2.57

0.27 -0.04 -027

-2.00

-8.03 -0.30 -1.65 0.09 -0.05

-0.25

- 1.42 - 0.83 - 1-40 - 0.72 0.04 - 0.87 - 127 0.04

0.05

0.06 -0.09 -0'37 0.13 -0.46 -0.19 0'01

0'00

0.06 -0-02 -0.02

0.05

-0.09 -0.05 0'00

-0.08

-0'52 0'01

-0.15

0.04 0.16

- 0'02

- 3'41

- 0.93

-0.12

0'22

0.15

0-05

001

0.26 - 3.09 - 5'35 -0-19

0'22

-1.80 -3'40

0.13 -0.74 - 1.63

0.06 -0.33 -0.76

0'02

0'12

0'10

0.28

0'28

0'22

0.13

- 0.91 - 1.46

-0.42 -0.93 -1.56 -4.64 - 3'97 -4.32 -2.42

-0.07 -0.46 -0.64 -1.68 0.03 -2.16 -0.89

-0.23 -0.24 -0.53 0.86 -1.04 -0.26

-426

0.29

-0'33

-0.25

0

-0.12

-0.10

0'02

- 0.86

7

0'00

-0.05

- 1.63 - 6'49

- 9'55 - 6.61

-00'02

-0.51

0'12

-8.25

0.08

-0.26 -0.03

0.18

-2.63

-0.04

0'02

0'00

0'00 0'02

-0.15 -0.27 0.03 0.04 0'01

-O.IO

-0.05 -0.10

0.5I

-0.38 -0.01 H G E 41

L. R. WEITKAMP AND

66

OTHERS

Table 2 (cont.) No. of Phase-known hetero- offspring? Chrornosome

Locus Pi GPT CA I1

parents* NFt 2F o 6 M 5 I5F 7

PTC Pb Pr(Db)

Lod scores$ for B of A

r

ZY~OUS

R I 2

9

3M

1

0

4F 3F

1

3

1

I

IF

0

0

IM

2

I

4F

0

0

0'I

0'2

-3.16 -4.55 - 8.98 0.09 - 3'98

- 2.03

- 0.98

-2.13

-0.61 - 1.91

- 2'32 - 0.72

- 1'47 - 0.44

- 0.44 - 3.09

-0.19 - 1-79

- 5.09 0.28

0.33 - 0.7I - 0.68

- 2.17

-0.4

0.3

0.05

-0.19 0'0I

- 0.70

- 0.45 - 0.03

-0.15

0.13

- 0.68

- 0.25

0.24 -0.18 - 0.29 - 0.08 0.07

- 0.06 - 0.08 - 0'02

- 0.25

- 0.06

0'1

I

0.06

* t

M = male; F = female; B = both. NR = non-recombinant ; R = recombinant. $ The lod scores include the data from phase known offspring; 0 = recombination frequency. 8 I n 8 male and 7 female Greek families Secretor was inforred from Lewis a type, i.e. Lewis a positive individuals were scored as Secretor negative and Lewis a negative as Secretor positive.

p loci*

Table 3. Linkage relationships of the hemoglobin a and Phaseknown offspring

Haemoglobin variants Source

4P

r-h7

Sex

Nli, It o I

Lod scores for 6' of A

r-

--_-7

0.05

0.1

0'2

0'3

0'4

-1'000

-0.699

-0.398

-0'222

- 0.097

0'279

0.255

G/thal

M

G/S

nf

I

0

6

o

z o o

I

o

G/D

M M M M

(1971) Rising et 02. (1974) G/S

M

4

0

Rucknagel & Rising

M

I

2

-1.770 -1'721

-0.754 -1.143

0.041 0.282 -0'592 -0.298

14

6

-5.167

-2.373

-0.182

0.506

F

I

2

- 1.721

-1.143

-0'592

-0.298

F F F

0

0

o

o

I

2

-0'392 0.008 0.197 0.148 0.258 0.215 0.134 0.065 -1.721 -1.143 -0'592 -0.298

o

o

4

6

Schwartz et al.

(1957) Scliwttrtz et nl.

(1957) Dherte et al. (1959) P/S Raper et al. (1960) G/C Lie-Injo et al. (1968) G/S ltothman & Ranney

G/S

2

0'204

0.146

0.079

1.532 1.673 1.225 0.877 0'475 0.010 0.070 0.062 -0.442 -0.188 0.032 -0.186 0.022 0.124 0.095 -2.000 - 1,398 -0.796 -0'444 -0.194

0.246 -0.115

(1975) Total

(MALE)

Scliwnrtz et ul.

G/t ha1

(1957) Brtldlcy et u Z . (1961) HOJS Pugh et nl. (1964) G/S Wong & IIuisman D/S

(1972) Rising et cil. (1974) Riic*lriu~gol & Rising

Total

F F

tlial/thal

F

(FEJIALII:)

Hdl-("ro,ggs et al. (1

G/S G/S

StII/B

Unspecifiod

0

0

6

10

o

o

20

16

0.535 -4.885

0.488 -0'115

0.048 0.0I 7 -0.115

0.465 0.318 0.170 0.049 -3.173 -1.571 -0.746 - 0.265

0.066 0'793 0.680 0,452 0.235 -7.133 -4.091 -1.654 -0.724 -0.315

- 1.182 -0.529 -0.216

- 0'053

- 14.245 -7.646 -2.365 -0.434

0'I 2 0

- 1.945

964) Total (BOTH)

*

The pedigrees were scored only for those sections in which both parents were tested.

Linkage relationships of haemoglobin loci with genetic marke? systems

67

On the basis of in situ mRNA hybridization, Price et al. (1972) have proposed that one of the haemoglobin linkage groups, most likely a,is on the long arm of chromosome no. 2 and the other is on the long arm of a B group chromosome. The interpretation of these mRNA-DNA hybridization data was met with scepticism (Bishop & Jones, 1972; Prensky & Holmquist, 1973). However, the original observations have been confirmed by in s i t u cDNA-DNA hybridization (Atwood et al. 1975). Further, using mRNA-DNA hybridization the mouse a and /? chain loci have been mapped t o chromosomes no. 11 and 7 - chromosomes already known t o carry the sites of a and /3 synthesis (Atwood et al. 1975, 1976). On the other hand, the interpretation which Anderson et al. (1976) place on their cDNA-DNA hybridization results in somatic cell hybrids leads to the tentative conclusion that the Hb, locus has been excluded from all but chromosomes no. 3, 12 and 14 and the Hb, locus from all but chromosomes no. 1, 7 and 15. The results of cDNA-DNA hybridization indicate that the a and /5’ genes are asyntenic, i.e. on different chromosomes (Deisseroth et al. 1976). I n some individuals there appear to be two a chain structural loci whereas in others there may be only one (cf. Rucknagel & Rising, 1975; Huisman & Miller, 1976). Some forms of a thalassaemia are produced by a deletion of the a chain structural gene (Ottolenghi et al. 1974; Taylor et al. 1974), but in other cases this may not be the situation (Kan et al. 1976). It seems probable, however, that all types of a chain variation are produced by genes in a single region. What effect, if any, a small deletion might have on recombination between the Hb, region and nearby marker genes is not clear. Further it is not known to what extent there may be population-specific effects on recombination frequency. Such effects may occur in the HLA :GLO linkage relationship (Weitkamp, 1976). Since by inspection of the data in Appendix 1 there were no significant differences among the results in the American black, Greek or Kar Kar Island population, whether segregating for a thalassaemia or a chain variants, these factors have been ignored in the summary presented in Table 1. To the data in this table may be added the results from the pedigree segregating for the a chain variant, Hopkins-2, described by Bradley et al. (1961). The combined data extend some regions in which Hb, maybe excluded: MN a t 8 < 26 yo, Hp, a t 0 c 15%, Le a t 0 < 5 % and P a t very close linkage. (Consistent with the protocol used in our own data, the linkage analysis in this pedigree was limited to the three female informative matings in which both spouses had been tested.) Based on an estimated total autosomal map length of 33 morgans (Renwick, 1971), the total data exclude the Hb, locus from approximately 7 % of the autosomal genome. Very close linkage of the /3, 6 and y loci may be inferred from study of the S/3 and y/5’ fusion haemoglobins (see Weatherall et al. 1974 for review). Both the loci determining /3 chain structural variants and /? thalassaemia are closely linked t o the 6 locus (Mishu & Nance, 1969; Weatherall et al. 1976). At least some varieties of hereditary persistence of foetal haemoglobin (HPFH’, are produced by deletion in the /3 and S regions (Huisman et al. 1975; Kan et al. 1975). It thus appears probable that all of the variants listed in Appendix 2 (haemoglobin S, C , B, /? thalassaernia, /?Sthalassaemia, and H P F H ) are controlled by loci in a single region. By inspection of the data, there is no significant heterogeneity in recombination frequency among the American black, American white or Greek populations for any of the marker loci tested. This is, of course, a necessary result if none of the marker loci are linked to the Hb, region. The major previous study on the linkage relationships of Hb, is that of Nance et al. (1970). 5-2

L. R. WEITKAMP AND

68

OTHERS

Other reports in which data concerning the Hb, linkage relationships with one or more marker loci may be found include Snyder et al. (1947), Snyder, Clarke & Moore (1949), Snyder (1949), Neel, Schull & Shapiro (1952), Ludwin, Limentani & Dameshek (1952), Dreyfuss (1955), Clarke et al. (1960), Bradley et al. (1961),Morganti et al. (1969), Weitkamp et al. (1972), Edwards (1974), Barbosa et al. (1975) and Palk et al. (1975). Many of these reports are concerned solely with the linkage relationships of the MN blood group locus with Hb,, an issue first raised by Snyder, Russel & Graham (1947). The subject received renewed interest (Weitkamp et al. 1972; Barbosa, Koury & Krieger, 1975) when it was thought there was an increased probability from other evidence that one of the Hb regions was on the long arm of chromosome no. 2 and the other on the long arm of chromosome no. 4 (Price et al. 1972) and that the 1MN locus was either on the long arm of chromosome no. 4 or no. 2 (German et al. 1969). It seems unlikely that the proposed assignment of MN to band 2q14 (German & Chaganti, 1973) can be correct (reviewed in Weit,kamp et al. 1976) and indeed the location of Hb, and Hb, is not certain. Elimination of five families in which the untested father was inferred to be a double heterozygote from the types of his spouse and offspring (a situation in which occult non-paternity of one child could give a false positive lod score) plus the addition of other completely tested families has reduced the peak lod score in our current data to the point where there is no indication of linkage between MN and either the Hb, or Hb, regions. In addition to the 34 marker systems included in this study results on the linkage relationships of Hb, with Inv, Ag, catalase and Diego have also been reported (Morganti et al. 1969; Nance et al. 1970). Inv and Ag may be excluded from linkage with Hb, a t 8 < 0.20 and catalase and Diego only for very close linkage. It is not clear to what extent the lod scores in Edwards (1974) and Falk et al. (1975) are derived from the same families (Falk et al. 1975). We have, therefore, considered only the results from Edwards where data on the same marker loci are reported by the two groups. Using the results in the above reports and calculations based on the pedigrees in Bradley et al. (1961) for Le and P, and in Barnicot, Garlick & Roberts (1960) for Tf, the exclusion of Hb, may be increased for HLA to 8 < 0.30 for Le, Kell and P to 8 < 0.25, for Gc to 8 < 0.20 and for Tf to 8 < 0.15. Based on the most likely estimates of map distance between the marker loci on chromosome 1 (fig. 3 in Hamerton, 1976), the Hb, locus may be excluded from the region encompassing PGD at the distal end of the short arm through Rh, PGM,, Amy and Fy. I n males the distance between PGD and Fy is approximately 106 centimorgans (cM) (fig. 3 in Hamerton, 1976). Since recombination is 50% more frequent in females for this region, the neuterized distance (male +female) is 132 cM. The Hb, region may be excluded by a distance of 25 cM from PGD and 28 cM from Fy. Thus, the Hb, region is excluded from 185 cM on chromosome no. 1 roughly two-thirds of the estimated 3 M length of this chromosome (Renwick, 1971). This calculation assumes, of course, that PGD itself is a t least 25 cM from the end of chromosome no. 1. If one takes into account the most likely distances between the known linkage groups (GLO:HLA, Kell: PTC, Lu: Se, lk:C3, Pi: Gm and Tf: El), then from the combined data we estimate, ignoring the obvious possibility that one or more of the marker loci may be quite near the end of a chromosome, that the Hb, locus has been excluded from about 11.6 M of a 33 M autosomal map.

-

N

Linkage relation.ships of haernoglobin loci with genetic marker systems

69

SUMMARY

An analysis of the linkage relationships of the Hb, and Hb, loci with 34 genetic marker systems is presented. No evidence of linkage of either haemoglobin locus with any of the marker loci was found. The Hb, locus may be excluded from approximately 7 yoand the Hb, locus from approximately one-third of the autosomal genome. This work was supported in part by N l H Contract NO1 HB 1-2404,Grant 1 ROl GM 1-19962,and Research Career Development Award 5 K O 4 H.D-50248 and by Grant 1-443 from the National Foundation. We thank Ms S. A. Guttormsen, E. Johnston, S. Hempfling, S. Libert, and L. Witinski for technical assistance.

REFERENCES

ANDERSON, W. F., DEISSEROTH, A. B., VELEZ,R., NIENHUIS,A. W., RUDDLE,F. H. & KUOHERLAPATI, R. S. (1976).A new technique for mapping the human hemoglobin genes. Human Gene Mapping 3. Birth Defects: Original Article Series 12 no. 7 pp. 367-71. ATWOOD, K.C., HENDERSON, A. S., KACIAN, D. & EICHER,E. M. (1975).On the feasibility of mapping low multiplicity genes by in SitU hybridization. Gene Mapping 2. Birth Defects: Original Article Series 11, no. 3, pp. 59-61. ATWOOD, K. C., Yu, M. T., EICHER, E. & HENDERSON, A. S. (1976). Feasibility tests for mapping lowmultiplicity genes by hybridization in situ. Human Gene Mapping 3. Birth Defects: Original Article Series 12, no. 7, pp. 372-5. BAGLIONI,C. (1962).The fusion of two peptide chains in hemoglobin Lepore and its interpretation as a genetic deletion. Proc. Natn. Acad. Sci. U.S.A. 48, 1880-6. BARBOSA, C. A. A., KOURY,W. H. & KRIEQER,H. (1975).Linkage data on MN and the H b p locus. Am . J . Hum. Genet. 27, 797-801. BARNICOT, N.A., GARLICK,J. P. & ROBERTS,D. F. (1960). Haptoglobin and transferrin inheritance in northern Nigerians. Ann. Hum. Genet., Lond. 24, 171-83. BISHOP, J. 0.& JONES, K. W. (1972). Chromosomal location of human haemoglobin structural genes. Nature, London 240, 149-50. BRADLEY, T. B. JR.,BOYER,S. H. & ALLEN,F. H. JR.(1961).Hopkins-2-hemoglobin: a revised pedigree with data on blood and serum groups. Bull. Johns Hopkins Hosp. 108, 75-79. CLARKE,C. A., DONOHOE, W. T. A., FINN, R., MCCONNELL,R . B., SHEPPARD, P. M., & NICOL,D. S. H. (1960).Data on linkage in man: ovalocytosis, sickling and the Rhesus blood group complex. Ann. Hum. Genet., Lond.24, 283-7. DEISSEROTH, A., VELEZ,R. & NIENHUIS,A. W. (1976). Hemoglobin synthesis in somatic cell hybrids: Independent segregation of the human alpha- and beta-globin genes. Science, N . Y . 191, 1262-4. DHERTE,P.,LEHMA”, H. & VANDEP~TTE, J. (1959).Hemoglobin P in a family in the Belgian Congo. Nature, Lond. 184, 1133-5. DREYFUSS, F. (1955).La thalassemie mineure (Cooley) dans trois familles juives orientales. J . Gdndt. hum. 4, 143-51, EDWARDS, J. H. (1974).Total lods for linkage analyzed by the New York Blood Center and Birmingham computer programs. Human Gene Mapping. Birth Defects: Original Article Series 10, no. 3, pp. 187-202. EFREMOV, G. D., HUISMAN, T. H. J . , SMITH,L. L., WILSON,J. B., KITCHENS, J. L., WRIGHTSTONE, R. N. & ADAMS, H. R. (1969).Hemoglobin Richmond, a human hemoglobin which forms asymmetric hybrids with other hemoglobins. J . Biol. Chem. 244, 6105-16. FALK, C. T., WALKER,M. E., MARTIN, M. D. & ALLEN,F. J. JR.(1975).Autosomal linkage in humans. Ser. Haemat. 8, 153-237. GERMAN,J., WALKER,M. E., STIEFEL,F. & ALLEN,F. H. JR.(1969).Autoradiographic studies of human chromosomes. 11. Data concerning the position of the MN locus. ?’ox Sang. 16, 130-45. GERMAN, J. & CHAGANTI, R. S. K. (1973).Mapping human autosomes: assignment of the niW locus to a specific segment in the long arm of chromosome no. 2. Science, N . Y . 182, 1261-2. HALL-CRAGGS, M., MARSDEN,P. D., RAPER,A. B., LEHMAN, H. & BEALE,D. (1964).Homozygous sickle-cell anemia arising from two different haemoglobins S. Interaction of haemoglobins S and Stanleyville 11. Brit. Med. J . ii, 87-9. HAMERTON, J. L. (1976).Report o f the committee on the genetic constitution of chromosomes 1 and 2. Human Gene Mapping 3. Birth Defects: Original Article Series 12, no. 7, pp. 7-23.

L. It. WEITIUMP AND

OTHERS

HUISMAN, T. H. J. & MILLER,A. (1976). H b Grady and a thalassemia: a contribution to the problem of Hb, structural loci in man. A m . J . Hum. Genet. 28, 363-9. HUISMAN, T. H. J., MILLER,A. & SCHBOEDER, W. A. (1975). A “ y type of the hereditary pcrsistcnce of fetal hemoglobin with J3 chain production in cis. A m . J . Hum. Genet. 27, 765-77. HUISMAN,T. H. J., WRIGHTSTONE, R. N., WILSON,J. B., SCHROEDER, W. A. & KENDALL, A. G. (1972). Hemoglobin Kenya, the product of fusion of y and J3 polypeptide chains. Arcla Biochem. Biophys. 153, 850-3.

KAN,Y .W., HOLLAND, J. P., DOZY,A. M., CHARACHE, S. & KAZAZIAN, H. H. (1975).Delotion of tlie P-globin structural gene in hereditary persistence of foetal haemoglobin. Nature, Lond. 258, 162-3. KAN,Y. W., DOZY,A. M., TRECARTIN, R. F. & TODD,D. (1976). Identification of a non-deletion type of a thalassemia-1 defoct. Abstracts. A m . SOC.Hematol. 19th Annual Meeting, Boston. KENDALL, A. G., OJWANG, P. J., SCHROEDER, W. A. & HUISMAN, T. H. J. (1973). Homoglobin Kenya, tho product of a 7-P fusion gene: Studies of the family. A m . J . Hum. Genet. 25, 548-63. LIE-INJO,L. E., WANG, A. C. & BURNETT, R. C. (1968). Anot,her family showing interaction of tlie gcncs for Hb G and Hb S. Acta Haemat. 40, 286-98. LUDWIN,I., LIMENTANT, D. & DAMESKEK, W. (1952). Linkage tests of Mediterranean anemia with blootl groups, blood types, IZh factors and eye color. Ant. J . Hum. Genet. 4, 182-93. MALAMOS, B., FESSAS, PH. & STAMATOYANNOPOULOS, G. (1962). Types of thalasscmia-trait, curriers as revealed by a study of their incidence in Greece. Br. J . Haemat. 8, 5-14. MISHU,M. K.& NANCE,W. E. (1969). Further evidence for close linkage of the H b b and Hbs loci in miin. J . illed. Genet. 6, 190-2. MORGANTI, G., BEOLCHINI, P. E. & GUALANDRJ, V. (1969). Lack of association and linkage botwecn P-thalasscmia and some serum protein systems (Gm, Inv, Hp, Gc and Ag). Humangenetik 7, 236-9. MORTON,N. E. (1955). Sequential tests for the detection of linkage. A m . J . Hum. Genet. 7, 277-318. NALBANDIAN, R. M., NICHOLS,B. M., CAMP,F. R. JR., LUSHER,J. M., CONTE, N. F., HENRY, 1%.L. & WOLF,P. L. (1971).Dithionite tubc test - a rapid, inosponsive technique for the detection of hcmoglobirr S and non-S sickling hemoglobin. Clin. Chem. 17, 1028-32. XANCE, W. E., CONNEALLY, M., KING,K. W., REED,T., SCHROEDER, J. & ROSE,8. (1970). Ccnctic linkagu andysis of human hemoglobin variants. A m . J . Hum. Genet. 22, 453-9. NEEL,J. V., SCHULL, W. J . & SHAPIRO, H. S. (1952). Absence of linkage between the genes responsible for the sicklirig phenomenon, the MN blood types, and the S-agglntinogen. A m . J . Hum. Genet. 4, 204-8. OTTOLENGHI, S., LANYON, W.G., PAUL, J.,WILLIAMSON, R., WEATHERALL, D. J . , CLEGG, J. R., PRITCHAKD, J.,POOTRAKUL, 8. & BOON,W. H. (1974).Tho severe form of a thalassomia is caused by a Iiaemoglobin gone deletion. Nature, Lond. 251, 389-91. PRENSKY, W. & HOLMQUIST, G. (1973). Chromosomal localization of human Iiaemoglobin st’ructuralgencs : techniques queried. Nature, Lond. 241, 44-5. PRICE, P. M., CONOVER, J. H. & H ~ S C H H O RK. N ,(1972). Chronioson~allocalization of human llaemoglobin structural genes. Nature, Lond. 237, 340-2. P U G H , 1%.l’.,MONICAL, T. v. & ~ ~ N N I C Hv. , (1964). Sickle cell anomin with two adult 1ioinogIot)iiwHb S and Hb G Philadelphia/S. Blood 23, 2 0 6 1 5 . RAPER,A. R., UAMMACK, D. B., HUEKNS,E. R. & SHOOTER, E. M. (1960). Four hacmoglobins in on0 individual. A stndy of the genetic interaction of H b C, and H b C. Brit. Med. J. ii, 1257-61. RENWICK, J . (1971). The m q p i n g of human chromosomes. A . Rev. Genet. 5 , 81-118. RISING,J. A,, SOUTTER, It. L., SPICER,S. J. (1974). Hemoglobin G-Philaclclphia/S: a family study of an inherited hybrid hemoglobin. A m . J . Clin. I’iLth. 61, 92-102. ROTIIMAN, M. & RANNEY,H. M. (1971). Doublc lietorozygosity for hemoglobin G (a368L y ~ / j % a ) nnd henwglobin D (a2*P2121ah). Blood 37, 177-83. ROWLEY, P. T., JACOBS, M., ROSECRANS, C., WEITKAMP, L. R. & DOHERTY, It. A. (1972). High rcsolution tinnlysis of licmoglobins : polyacrylarnido isoclectric focusing. Biochem. A f ell. 6, 663MiO. RUCKNAGEL, D. L. & NEEL,J. V. (1961). The hemoglobinopathies. Prog. N e d . (;en&. 1, 158-360. RUCKNAGEL, D. L. & RISJNG,J. A. (1975). A hoterozygote for Hbf, HbbCand H t ~ a a r ’ h i ~ a ~ e i pinh ~1% ~ fmnily presenting evidenco for heterogeneity of homoglobin alpha chain loci. A m . J . Jferl. 59, 53-60. SCHWARTZ, H. C., SPAET,T. H., ZUICI.ZElE, W. W., NEEL,J. V., RODINSON, A. R. & KATJBMAN, 8. F. (1967). Combinations of hemoglobin G, S and thalassemia occurring in ono family. Blood 12, 238-50. SCHWARTZ, E. (1976). Multiple loci for human globin genes. A m . J . Human Geuet. 28, 423-5. SHOWS, T. B. (1975). Gene markers for mapping the liunian genoino. Cytogeib. Cell Gcnct. 14, 199 207. SNYDER, L. H. (1949). Studies in human inheritanco. XXXV. The linkage relationship of sickle cc.11 nriuinia. Hereditas. Supp. Proc. V I I I Int. Congr. Genet. pp. 4 4 6 5 0 . SNYDER, L. H., CLARKE:, H. & MOORE,C. V. (1949). Studies in human inheritance. XXXIV. Furtlior tlirta on the linkage of the genes for sickle cells and the M-N blood types. Ohio J . Sci. 49, 32-3. SNYDER, L. H., HUSSEL,H. & GRAHAM, E. B. (1947). Linlcage between the genes for sicklo cells iind the M-N blood types. Science, N . Y . 106, 347-8.

Linkage ?.elationshipsof haernoglobin loci with genetic marker systems

71

STAMATOYANNOPOULOS, G. & NUTE,P. E. (1974). Genetic control of hoomoglobins. Clinics in Haem. 3, 251-87.

TAYLOR, J. M., DOZY,A,, KAN,Y .W., VARMUS,H. E., LIE-INJO,L. E., GANESAN, J. & TODD,D. (1974). Genetic lesion in homozygous a thalrtssomia (hydrops fetalis) . Nature, Lond. 251, 392-3. WEATHERALL, D. J., CLEGG,J. B., MILNER,P. F., MARSH,G. W., BOLTON, F. G. & SERJEANT, G. R. (1976). Linkage relationships between p- and 8-structuralloci and African forms of thalassemia. J . Med. Genet. 13, 20-6. WEATHERALL, D. J., PEMBREY, M. E. & PRITCHARD, J. (1974). Fetal haemoglobin. Clinics i n Haem. 3, 467-508.

WEITKAMP, L. R. (1976). Linkage of GLO with HLA and Bf. Effect of population and sex on recombination frequency. T i s . Antigens 7 , 273-9. WEITKAMP, L. R., ADAMS,M. S. & ROWLEY, P. T. (1972). Linkage between the MN- and Hb p-loci Hum. Heredity 22, 566-72. WEITKAMP, L. R., FERGUSON-SMITH, M. A., GUTTORMSEN,S. A., HUNTZINGER, R. S., CHAGANTI,R. S. K., GERMAN, J. & SCHANFIELD, M. S. (1978). The linkage relationshipsof marker sites on chiomosomes No. 2 and 10. (In manuscript.) WOKEN,D. ( 1 970). Description of MOSM :Program for Genetic Linkage Analysis. Oslo :Norwegian Computing Center. WONG,S. C. & HUISMAN, T. H. J. (1972).Further evidence for non-linkage of Hb, and Hba structural loci in man. Clin Chem. Actu 38, 473-4.

Note added in proof Since preparation of this manuscript Deisseroth et al. (1977), using annealing of a globin cDNA to DNA extracted from mouse/human somatic cell hybrids containing a variable number of human chromosomes, have reported that the Hb, locus is on chromosome 16. Among the loci we tested for linkage, Hp, is the only one assigned t o chromosome 16. Our data, principally derived from a-thalassemia in Greeks, plus the data from the Hopkins-2 variant described by Bradley et al. (1961) exclude linkage of Hb, with Hp, a t 13 < 0-15. DEISXEROTH, A., NIENEUIS,A., RUDDLE,F., LAWRENCE, J., CREEGAN, R . , VELEZ,R., TURNER,P., KUCFIERLAPATI, R. & ANDERSON,W. F. (1977). Localization of the human alpha globin gene to chromosome 16. Clin. Res. 25, 3 2 2 ~ .

L. R. WEITKAMP AND

72

OTHERS

Appendix 1. Genetic linkage relationships of the haernoglobin a locus

Marker , - - -A- (

Chromosome I

Locus PGD

Rh PGM,

6

HLA

Haemoglobin variant

PhaseNo. known of ORhetoro- spring Popu- zygous 6 lation parents NR R

thal. Groek white thal. Greek white thal. Greek white 'D' American black thal. Greek white

IF

0

0

Lod score for 0 -7

0'1

0'2

0.3

0.4

- 0.72 I

- 0.444

- 0.194

- 0.076

- 0.01 8

1'347

1.185 - 0.652 - 0.444 - 0.652

04335 -0.130 -0,194 -0.130 0.214 -0.582 - 0.388 0.161

0.468 0.006 -0.076 0.006 0.216 - 0.227 -0.151 0'1 19

0.143

0'05

2M

0

0

5F IM 5F IM IF

0

0

0

0

0

0

3

I

0

0

IM

0

0

4F

0

0

thal. Greek white J Kar Kar Islander

3M 5F IF

0

0

0

0

0

0

0.258

'D '

IM IF

0

4

0

0

IM 2F

0

0

0

0

-4.000 - 1.185 0.533 -1'442

IF

0

I

J

American black Kar Ka,r Islander American black

- 1'372 - 0.721

- 1.372 -0.163 - 2.164 - 1.442 -0.188

0.066 - 1'331 - 0.887 0.046

-0.650

-0.207

- 3'071 - 1.753

0.134

0.064

-2.796 -0.673 0.465 -0.887 - 0.699

-1.592 -0.254 0.318 -0.388 - 0.398

-0.087 0.170 -0.151 - 0.222

0.049 -0.036 - 0.097

-2.627 - 3.208 -0.163 0.279

- 1.561 - 1.857

-0.642 -0.707 0.214 0.204

-0.238 - 0.232 0.216 0.146

-0.054 - 0.045 0.140 0.079

-4'327

-2.662

- 1.163 - 0'I 2 I

-0.454

-0.106

thal. Greek white ' D ' American black

3M 6F

0

0

0

0

IM

3 I

I 0

thal. Greek white 'D ' American black Unknown Lu thal. Greek white So (Le a) thal. Greek white Le 'D ' American black K thal. Greek white Js 'D ' American black MNSs thal. Greek white 'D' American black J Kar Kar Islander P Greek white ' D ' American black 'D' American Jk black thal. Greek Gm white thal. Greek Gc white thel. Greek E2 white

4M 4F IF

0

0

0

0

0

0

IM IF 2F

0

-0'721 -0.721

0

0 0 0

IF

0

IF

0

Hpa

IM IF 2M 6F IM IF IM 2F 2M 2F IF

0

0.035 0'039 0.031 - 0.039 0.017

0.215

ABO

16

- 0'053

- 0.208

'D '

IF

0.083

0'012 0.I 40

- 0.65 I

GLO 9

0.064

0'012

-0.018

- 1.000

0.066 0.255

- 0.927 - 0.458 0.258

0.215

0.134

-0.888

- 0'022

0.064

-0.388 -0.018

- 0.001 0.017

-0.194 -0.194 - 0'249

-0.076 -0.076

- 0.858

-0'444 - 0.548

- 0.100

-0.018 -0.018 - 0.029

0

- 1.185

-0.673

-0.254

-0.087

-0.018

0

-0.186

0

0

- 1'442

0

0

0.282

0

0

0

0

2

- 2,884 - 1.834 - 1'442 - 1'000 - 0.721

-0.444

0.022

- 0.887

0.124

0.095

- 0.388

-0.151 0.077 - 0.301

0.240 - 1'774

0.154 - 0.775 - 0.880 -0.190 - 0.888 - 0.388 - 0.699 - 0.398 - 0'444 -0.194 - 0.476 -00'212 -0.169 - 0'073 - 0.508 - 0229 0.240 0.154

0

2 I

0

0

0 0

0 0

0

0

- 0.803

0

0

0.282

IF

0

0

IF

0

0

2F

0

0

-0'907

-0.422

-0,070

IM

0

0

IF

0

0

-0.721 0.533

-0'444 0.465

-0.194 0.318

- 0.762

- 0.257

- 0.050

-0.152 - 0'222 - 0.076

0.031 -0.035 0'02 I

- 0.07 I 0'012

- 0.036 - 0.097

-0.018

- 0.084

- 0'020 - 0.027 - 0.006 - 0.091 - 0'022

0'077

0'02 I

0.019

0.014

-0.076 0.170

-0'018

0.049

Linkage relationships of haemoglobin loci with genetic marker systems

73

Appendix 2. Genetic linkuge relationships of the haemoglobin /3 and S loci No. of Marker A -,rHaemoheteroChromoglobin zygous some Locus variant Population parents I PGD thal. Greek 3M white 2F s, c American 8F black PGM, thal. Greek IZM white IIF American zF white s, c American 8M 12F. black American Amy SC 2M black 2F Fy thal. Greek 6M white 16F American IM white IF S Greek I F white S, C, B, American 5M black 8F 2

AcP,

thal.

S S, C, B,

Gt

Greek white American white American white American black

Phaseknown offspring h NR R

o o

o o

II

II

o o

o o

o

o

I

8 14

10 I

o o o o o o

2 2

o o o

- 0.780

- 0.179 0.055

- 5296 - 2,919 - 10.568 - 4'947 - 1.183 - 0.662 - I ~842 - 0.990 - 2.103 - 0'955 - 2.369 - 0'773 0.215 0.134

- I '463 - 0.656 -2.185

- 0'379

- 0.690 -0.177

-0.519 -0.381 - 0.209 0.064 0.146 -0.024

0.017 0.079 -0.006

-00'212

- 0.090 - 0.032

2

-1.998

-0.480

-0.099

o

-0.070 0.227

o

o

-0229 -0.444

0.289 -0.194 -0.060 -0,194

I

I

o

9M F

11

8

9 4

-7.839 -8.560

-4'599 -3-888

I

3

-5.091

o

I

M

o

o

I

2

0.435 -0.m

0.280

0.173 -0.076

0.083 -0.018

-0.011

-00'001

-0.018

-1.784 -0.412

-0.552

-0.037 0-394

-3.187

- 1.498

-0.693

-0.464

-0.229

-0.060

-0.011 -0.001

-0.444 -1.143 - 0.229 -1-324 -2.166 0'559 -0.444

-0.194 -0'592 - 0.060 -0.384 -0.682 0'254 -0.194 -0.855 -2.665 -1.962

-0.076 -0.298

-1.906

IB 7M 7F I B

o 4

o

o

o o 3 8 8

2

-1.811 -0.156

-0.076

-0.721 -1.721 - 0.464 -2.556 -3'976 0.730 -0.721 -3.350 - 11.860 -17.378

I

- 0.001

o 3

o

20

- 0.01I

-0.302

0.255

-4.134 -4275 0.517 -0.721 -0.464 -0.721

12M 13 F IB I M 3F I M

0.076

-0.055

American white

C

- 2.059 -0.278 - 0.060

-0.932

GLO

ADA

- 4.895 - 1.688 - 0.229

-0250

6

20

-0.036 -0.036 -0.179

-0.702 -2.140

o

P e p A S,C

-0.163 -0.151 -0.791

- 1.789 -4.747

M

18

-0.448 -0.388 -2.119

-3.091 -7.718

I

S, C, B,

-1.116 -0.887 -5.124

o 3

American black

tlial.

0.258

0.4

I

o

PGMa C

HLA (Bf)

- 4'377

\

0.3

-0.104

4

American black

- 9'257

- 16.724 - 1.464 - 2.721 - 3.308

0'2

o

3F

s, c

- 1.906

- 1'443 - 8.603

0' I

0,204 -0.056

American black

thal.

A

0.05

0.279 -0.137

I

C

201-3

Lod score for 0 f-

0.509

-0.249

-0.018 -0.115

- 0.01I

- o'ooo

-0.079 -0.109 0.062 -0.076 -0.354 -0.945 0'132

-0.006 0.068 0.004

American white American black

M

o

4F 16M 30F

2

American black

2M 4F

o

o 3

-5.812

-1.116 -3.631

-0.448

I

-1.692

-0.163 -0.769

-0.036 -0.266

American black

I

M

I

2

-1.721

-1.143

-0.592

-0298

-0.115

I

7 13

-2.008

-6.860 -8.657

-0.018 -0.119

-0.234 0.386

L. R. WEITKAMP AND

74

OTHERS

Appendix 2 (cont.) No. Marker

*,-., Cliromosome Locus UnLu known

of OffHaemohetero- spring globin zygous A variant Population parents NR R

thal.

Greek white

S Se thal. (Le a)

S So

thal.

s,c Le

Phaseknown

thal.

S,c

American black Greek white Greek white American white American black Greek white American black

thal.

C Kp

thal.

Js

S,C

Greek white American black American white American black

MNSs tlial.

P

Jk

Tf

Greek white American wliite Greek S white American white S, C, B, American black tlial. Greek white American white S Greek white 6 American black thal. American whito s, c American black thal.

American white

0'I

0'2

-0.229 -0.887

-0.060 -0.388 0'134

2M IF IF

0

0

0 0

0

- 0.464 - 1'442

0

0.258

0.215

2M

I

0

- 0'442

-0.188

7M

0

0

- 5'785

-3.555

5F IM 2F IF

0 0

0 0

0

2

0

0

2

5

I 2

4 M 9F

- 1.648 -0.901 0.814 -2'137 0.104

- 2.245 - 1'938 - 0.426

0.010

0.3

0.4

-0.011 -00'001

-0.151

0.064 0.070

-0.035

0.017 0*062

- 1.546 -0.601 -0.140

-0.098 -0.019 0.298 0.094 - 1.502 - 0.852 - 0.468 - 0.200 0.023 0.006 0.049 0.084 0.720

- 1'221 - 0.653

-0.199 0.084 - 3.486

-0.315

0.517

- 0.388 - 0.070

0.286 - 0'043 0'049 - 1'354 - 0764 0.042 0.318 0.124 -0.194

0'677 - 0.004

0.03 I 0.260

2M IF

0

0

0

0

M

0

0

16F

4

4

IM

0

0

2F IF

0

0

0

0

0.104 - 5'958 - 4'771 0'179 0'533 -0.186 - 0.72I

IF

0

0

- 1'442

-0.887 -0.388

-0.151

-0.035

- 3'235

- I ,756 -0.617 - 3'275 - 1.188

- 0'222

- 0.081 - 0'073

11

IB K

0.05

8M

2

3

F IB 21 M 18F IM 5F 5F

0

0

0

0

IM

10

- 5.841 0.321

- 12.581 - 9'417 - 0.72I

0

0

0

0

4 I

I I

- 0.348 - 3'349

0

0

0.258

-2.512 0'120

0.465 0'022

- 0.444

0.023

0'001

0.006

- 0'457 - 0.091 - 0.208 - 0.060 0'00I 0.009 0.170 0,049 0.03I 0.095 - 0.076 -0.018

- 0'379

0.018 0'001 0.085 0.226 0.01 8 -6.719 - 2.046 - 0.395 - 5,269 - 1.866 -0.550 - 0.090 - 0'444 -0.194 - 0.076 -0.018 0.219 0.351 0.358 0.093 - 2'004 - 0.836 -0.314 - 0.072 0.215

0'134

0.108 - 3'975 -1.587 - 24.264 - 14.296 -5.681 - 3'399 -2'051 -0.871

0,064

0.017

12M

5

27 F 7M

20

3 19

0

0

10F

0

I

3F

0

0

- 3.296 - 1'235

0.275 -0.318 -0'077 -2.061 -0.928 -0.392 -0.119 -0.719 -0.289 -0.106 -0.024

IF

0

0

-0'137

- 0.104 - 0.056 - 0.024

IB

- 0.006

0.009

0'001

- 1.885

- 1.076 -0.377 -0.081

0.026

-0.861 -0.245 -2.333 -0.552 0'149 0'053 0.487 0'442

-0.017 -0.038

0.043 0.129

0'011

0'001

0.265

0.081

0.179

2F

2

I

3M F IB 2F

I 2

0 0

- I .627 - 4.604

0

0

0.219 0'349

10

0.462

-2.053 -0'333

0'120

0'042

Linkage relationships of hemoglobin loci with genetic ma.rker systems

75

Appendix 2 (cont.) PhaseNo. known Marker of Off, -A- f Haemohetero- spring Chromoglobin zygous 7 4 some Locus variant Population parents NR R

s,c high F

El

thal

.

S

E, CP Gc

thal.

s, c thal.

S

s,c c3

thal.

R

s, c PTC

thal.

GPT

thal.

s,c CAII

s, c

Pi

S

Pb

C

Pr (Db)

thal .

S, B,

American black Greek white Greek white American black Greek white American black Greek white American white Groek white American black American whito American white American black American xzhite American white American black American black American black American black American white American black

Lod score for 6, 0.05

0'1

0'2

0.3

0.4

-1.183

-0'595

-0.229

2F

I

3

- 3'442

IM

0

0

0.258

0.215

0.134

- 1.906 -0.186 - 3'442

- 1.116

-0.448 0.124 - 1.183

2M

0

0

2F 2M

0

0

2

4

2M

0

0

3F 2M 3F 7M 14F 3F

0

0

IM 4F 6M 11 F 2F

0.276

-2.285

0.022

-2.285

0.064

0.017

-0.163 -0.036 o*ogg 0.031 -0.595 -0.229

0.275 0.221 0.130 -0.422 -0.070 0.019 - 0.928 - 0.458 - 0'233 - 1.560 -0.642 -0.238 - 2.426 - 0-go5 - 0.295 -0.824 0.275 0.374 -1.116 -0.448 -0.163

0.170 -0.036

- 0.464 - 1.628 - 3'534

I

2

-0 9 0 7 - 1.464

0

0

- 2.627

0

0

I

0

0

0

- 4.256 - 2'577 - 1.906

0.039 0.014 -0.097 -0.053

- 0.058

0

0

I

0

0

0

- 3'438 - 1.185

- 0.299 -0.861 -1.982 - 1.167 - 0.673

2M

I

0

- 0'442

-0.188

3M 3 F 3 F

3

7 0

I

I

-6.164 - 3'070 -2.319

-4'127 -2'173 - 1.748 -0.632 - I -466 - 0.678

-1.114 -0.168 - 0.289

-0.441

I

4 F

I

3

- 3'442

-2.285

-0.595

-0.229

6M

5 6

2

-4.554

-2.125

-0.611

6

- 5'534

- 2.804

- 0.726

- 0.087 - 0.019

0.328 -0.708 - 0.980

0'241 -0.179 - 0.450

-0.055 - 0.15 I

11 F

0

0

I

0

- 0.060 -0.244 -0.711 0'444 - 0.254

- 0'01 1

- 0'001

-0'017 -0.219 0.661 - 0.087

-0'043 -0.040 0'329 - 0.018

0.010

0.070

0.062

-1.183

-0.027

0.008

- 0.083

0.131

I

0

0.093

I

3

- 3'977

0

I

-3'163

0.277 -2.171 - 2-03I

IF

0

0

- 0.72 I

- 0.444

- 0' I94

- 0.076

- 0.018

IF

0

0

0.258

0.215

0,134

0.064

0.017

IM 3F

2

I

0

0

- 0'442 - 3.348

-0.189 - 2.004

- 0.836

3M 4F 2F

0.010

0.070 - 0.3 14

0.111

0.061

- 0.072