3654 Nucleic Acids Research, Vol. 18, No. 12
Nucleotide sequence of an actin cDNA gene from Aplysia californica L.DesGroseillers*, D.Auclair and L.Wickham Department of Biochemistry, University of Montreal, P.O. Box 6128, Station A, Montreal, Canada H3C 3J7 EMBL accession no. X52868
Submitted May 7, 1990 We present the complete nucleotide sequence of an Aplysia califomica actin cDNA gene isolated from an abdominal ganglion cDNA library (Miles,M. and Scheller,R.H., Stanford University; unpublished) (Fig. 1). Using PCR technology, we showed that this actin cDNA is expressed in the sheath surrounding the neurons (probably in the muscular cells of the sheath), while another homologous actin gene is expressed in the neurons (Fig. 2).
ACKNOWLEDGEMENTS We thank Mme T.Buy for technical assistance. This work was supported by Medical Research Council of Canada and Fonds de la Recherche en Sante du Quebec grants to L.D.G. L.D.G. is also a scholar from MRC.
M C D D E V A A L V V D N G S G M C K A G 1 acgaactttgttcactttccaaccttccag tccgttactcacacacaaactcaatATGTGCGACGACGAGGTTGCTGCTCTTGTTGTGGACAATGGCTCCGGCATGTGCAAGGCCGGC R P H Q G V M V G M G Q K D S Y V G D E A Q S F A G D D A P R A V F P S I V G R 121 TTCGCCGGTGACGACGCTCCCAGGGCCGTCTTCCCCTCCATCGTCGGACGCCCCAGGCATCAGGGTGTCATGGTTGGTATGGGCCAGAAGGACAGCTACGTCGGTGATGAGGCTCAGTCC. K R G I L T L K Y P I E H G I V T N W D D M E K I W H H T F Y N E L R V P P E E 241 MQAGAGGTATCCTCACCCTGAAGTAC CCC ATCGAG CACGGCATCGTCAC CAACTGGGATGACATGGAGAAGATC TGGCATCACAC CTTCTACAATGAG CTG CGTGTT CCC CCAGAGGAG H P V L L T E A P L N P K A N R E K M T Q I M F E T F N A P A M Y V A I Q A V L 361 CAC CCCGTCCTG CT CACTGAGGC CCCCCT CAACCC CAAGGCCAACAGAGAGAAGATGAC CCAGATCATGTTCGAGAC CTTCAACGC CCCCGC CATGTACGTTGCCATCCAGG CCGTGCTC S L Y A S G R T T G I V L D S G D G V T H T V P I Y E G Y A L P H A I L R L D L 481 T CC CTGTACGC CTC CGGACGTACCACAGGTATCGTG CTTGACT CTGG CGATGGTGTCACC CACACCGT CCC CATCTACGAGGGTTACG CTCTGC C CCACGC CATCCT CCGTCTGGACTTG A G R D L T D Y L S K I L T E R' G Y S F T T T A E R E I v R D I K E X L C Y V A 601 GCTGGCCGTGACTTGACCGACTACCTGAGTAAGATCCTGACCGAGCGTGGCTACTCTTTCACCACCACCGCCGAGAGAGAGATC,GTG.CGTGACATCAAGGAGsGCTGTGCTACGTCGCC L D F E Q E M A T A A S S S S L E K S Y E L P D G Q V I T I G N E R F R C P E S 721 CTGGACTTCGAGCAGGAGATGGCCACCGCCGCCTCCTCCTCCTCCCTGGAAAAGAGCTACGAGCTTCCCGACGGACAGGTCATCACCATCGGAAACGAGCGCTTCAGGTGCCCCGAGTCT L F Q P I L L G M E S A G I H E T T Y N S I M K C D V D I R K D L Y A N T V L S 841 CTCTTCCAGCCAATCCTTCTTGC7,GATGGAATCITGCTGGTATCCACGAGACCACCTACAACTCCATCATGAAGTGCGACGTGGACATCCGTAAGGATCTGTACGCCAACCCGTCCTGTCC G G T T M F P G I A D R M Q K E I T S L A P S T M K I K I I A P P E R K Y S V W 961 GGAGGCACCACCATGTTCCCTGGTATTGCTGACCGCATGCAGAAGGAGATCACCTCTCTGGCTCCCAGCACCATGAAGATCAAGATCATCGCTCCCCCTIGAGCGCAAATACTCCGTATCGG I G G S I L A S L S T F Q Q M W I S X Q E Y D E S G P S I V H R K C F * 1081 ATCGGTGGCTCCATCCTGGCTTCTCTCTCCACCTTCCAGCAGATGTGGATCTCCAAGCAGGAGTACGATGAGTCCGGCCCCTCCATCGTGCACCGCAAGTGCTTCTAAaqcqctcqcccq 1201 1321 1441 1561 1681 1801 1921 ccaaaqattqqccagtcqaaqtgtcaaaqtttttqgtaacaacaacaccqcqtcaaqttatacaaqcaaaqaatacaatqaagtaaqtctggtcscctactatgtaacatcaaqaacacq 2041 2161 2281 aaattactqtaqaccttcqaaaqqctgaataaaatgtgttqaattaaqtg(poly A)42
cactccaacaacatccaacaacatcaqaaqaaqaacaaatcqqccactcqcqaqcatttaqacctatcqctcqaqgacattqacaataacatqacatttgtcaaaaqaaqcaacaactca acaaqtcqactqaaacttataacaacaacagtaaqaacqagtqqctqaaaaqacattqaaaqaacquaaaaaaaaqaacaaaaaaqatttaattgstatqaacatcttcqctacaatttcc aaccctacttqaaqtcttaaaattqaaaaqcgtatgttactttaatcttacaqaaacctgtatqggaqaqggaqcatactcaaatttqgactttaaqatttttttgtaqqggaaaaaaact ttqcqataaaaagtttatctttttatcccattqcaqcctttqatttttgtttcgttatctaatatttcatttaactcataagtcqacaactatcatctaactgttatactaqqatqaaaq atqqqaqqcqaacattacttatcqaaqgctacaqcttcactcaqgaaaacaaaatgttatcqaaaaqaaqtaacattttcttttctacacctaqgstcttqccttqaqcqaqtaatccaa tctccatqacaagcaaacqaaaqatttgtgtaaattttcctaaqtttttttttataaaaaquaaaatcctqaaaaacctqcqcgtacgaaaqtqgtgtgtcaatqcaqtttgtgtqaacgt taacqactaattctqaactccaacaaaqaatgstcaaaqacaatatqgtcccttcqctttqatqccaaqactqqacqcagacataaatgttqacqttttqcaaatctacctggcqcgtac cttatttqtqcqacaaactcccatcctqccgttqctataqaaacqattqgtcgtttccatgqtaqcqgtaaatcqtcatcaagcttaqctatctqctttqqtttctqtacatqcattatc
Figure 1. Actin gene sequence. The positions of the 3 primers are underlined in the sequence.
0LIGCO T
5 SUE S
S N
S N M
OR F
.L ZZ}A-
-.-_.
Figure 2. PCR amplification of reverse transcribed mRNA isolated from the sheath (S) or from the neurons (N), using either two primers derived from the coding region (primers 2+3) or one primer from the coding region and one from a specific 5' untranslated region (primers 1+3). M: marker. *
To whom
correspondence
should be addressed
Nucleotide sequence of an actin cDNA gene from Aplysia californica L.DesGroseillers*, D.Auclair and L.Wickham Department of Biochemistry, University of Montreal, P.O. Box 6128, Station A, Montreal, Canada H3C 3J7 EMBL accession no. X52868
Submitted May 7, 1990 We present the complete nucleotide sequence of an Aplysia califomica actin cDNA gene isolated from an abdominal ganglion cDNA library (Miles,M. and Scheller,R.H., Stanford University; unpublished) (Fig. 1). Using PCR technology, we showed that this actin cDNA is expressed in the sheath surrounding the neurons (probably in the muscular cells of the sheath), while another homologous actin gene is expressed in the neurons (Fig. 2).
ACKNOWLEDGEMENTS We thank Mme T.Buy for technical assistance. This work was supported by Medical Research Council of Canada and Fonds de la Recherche en Sante du Quebec grants to L.D.G. L.D.G. is also a scholar from MRC.
M C D D E V A A L V V D N G S G M C K A G 1 acgaactttgttcactttccaaccttccag tccgttactcacacacaaactcaatATGTGCGACGACGAGGTTGCTGCTCTTGTTGTGGACAATGGCTCCGGCATGTGCAAGGCCGGC R P H Q G V M V G M G Q K D S Y V G D E A Q S F A G D D A P R A V F P S I V G R 121 TTCGCCGGTGACGACGCTCCCAGGGCCGTCTTCCCCTCCATCGTCGGACGCCCCAGGCATCAGGGTGTCATGGTTGGTATGGGCCAGAAGGACAGCTACGTCGGTGATGAGGCTCAGTCC. K R G I L T L K Y P I E H G I V T N W D D M E K I W H H T F Y N E L R V P P E E 241 MQAGAGGTATCCTCACCCTGAAGTAC CCC ATCGAG CACGGCATCGTCAC CAACTGGGATGACATGGAGAAGATC TGGCATCACAC CTTCTACAATGAG CTG CGTGTT CCC CCAGAGGAG H P V L L T E A P L N P K A N R E K M T Q I M F E T F N A P A M Y V A I Q A V L 361 CAC CCCGTCCTG CT CACTGAGGC CCCCCT CAACCC CAAGGCCAACAGAGAGAAGATGAC CCAGATCATGTTCGAGAC CTTCAACGC CCCCGC CATGTACGTTGCCATCCAGG CCGTGCTC S L Y A S G R T T G I V L D S G D G V T H T V P I Y E G Y A L P H A I L R L D L 481 T CC CTGTACGC CTC CGGACGTACCACAGGTATCGTG CTTGACT CTGG CGATGGTGTCACC CACACCGT CCC CATCTACGAGGGTTACG CTCTGC C CCACGC CATCCT CCGTCTGGACTTG A G R D L T D Y L S K I L T E R' G Y S F T T T A E R E I v R D I K E X L C Y V A 601 GCTGGCCGTGACTTGACCGACTACCTGAGTAAGATCCTGACCGAGCGTGGCTACTCTTTCACCACCACCGCCGAGAGAGAGATC,GTG.CGTGACATCAAGGAGsGCTGTGCTACGTCGCC L D F E Q E M A T A A S S S S L E K S Y E L P D G Q V I T I G N E R F R C P E S 721 CTGGACTTCGAGCAGGAGATGGCCACCGCCGCCTCCTCCTCCTCCCTGGAAAAGAGCTACGAGCTTCCCGACGGACAGGTCATCACCATCGGAAACGAGCGCTTCAGGTGCCCCGAGTCT L F Q P I L L G M E S A G I H E T T Y N S I M K C D V D I R K D L Y A N T V L S 841 CTCTTCCAGCCAATCCTTCTTGC7,GATGGAATCITGCTGGTATCCACGAGACCACCTACAACTCCATCATGAAGTGCGACGTGGACATCCGTAAGGATCTGTACGCCAACCCGTCCTGTCC G G T T M F P G I A D R M Q K E I T S L A P S T M K I K I I A P P E R K Y S V W 961 GGAGGCACCACCATGTTCCCTGGTATTGCTGACCGCATGCAGAAGGAGATCACCTCTCTGGCTCCCAGCACCATGAAGATCAAGATCATCGCTCCCCCTIGAGCGCAAATACTCCGTATCGG I G G S I L A S L S T F Q Q M W I S X Q E Y D E S G P S I V H R K C F * 1081 ATCGGTGGCTCCATCCTGGCTTCTCTCTCCACCTTCCAGCAGATGTGGATCTCCAAGCAGGAGTACGATGAGTCCGGCCCCTCCATCGTGCACCGCAAGTGCTTCTAAaqcqctcqcccq 1201 1321 1441 1561 1681 1801 1921 ccaaaqattqqccagtcqaaqtgtcaaaqtttttqgtaacaacaacaccqcqtcaaqttatacaaqcaaaqaatacaatqaagtaaqtctggtcscctactatgtaacatcaaqaacacq 2041 2161 2281 aaattactqtaqaccttcqaaaqqctgaataaaatgtgttqaattaaqtg(poly A)42
cactccaacaacatccaacaacatcaqaaqaaqaacaaatcqqccactcqcqaqcatttaqacctatcqctcqaqgacattqacaataacatqacatttgtcaaaaqaaqcaacaactca acaaqtcqactqaaacttataacaacaacagtaaqaacqagtqqctqaaaaqacattqaaaqaacquaaaaaaaaqaacaaaaaaqatttaattgstatqaacatcttcqctacaatttcc aaccctacttqaaqtcttaaaattqaaaaqcgtatgttactttaatcttacaqaaacctgtatqggaqaqggaqcatactcaaatttqgactttaaqatttttttgtaqqggaaaaaaact ttqcqataaaaagtttatctttttatcccattqcaqcctttqatttttgtttcgttatctaatatttcatttaactcataagtcqacaactatcatctaactgttatactaqqatqaaaq atqqqaqqcqaacattacttatcqaaqgctacaqcttcactcaqgaaaacaaaatgttatcqaaaaqaaqtaacattttcttttctacacctaqgstcttqccttqaqcqaqtaatccaa tctccatqacaagcaaacqaaaqatttgtgtaaattttcctaaqtttttttttataaaaaquaaaatcctqaaaaacctqcqcgtacgaaaqtqgtgtgtcaatqcaqtttgtgtqaacgt taacqactaattctqaactccaacaaaqaatgstcaaaqacaatatqgtcccttcqctttqatqccaaqactqqacqcagacataaatgttqacqttttqcaaatctacctggcqcgtac cttatttqtqcqacaaactcccatcctqccgttqctataqaaacqattqgtcgtttccatgqtaqcqgtaaatcqtcatcaagcttaqctatctqctttqqtttctqtacatqcattatc
Figure 1. Actin gene sequence. The positions of the 3 primers are underlined in the sequence.
0LIGCO T
5 SUE S
S N
S N M
OR F
.L ZZ}A-
-.-_.
Figure 2. PCR amplification of reverse transcribed mRNA isolated from the sheath (S) or from the neurons (N), using either two primers derived from the coding region (primers 2+3) or one primer from the coding region and one from a specific 5' untranslated region (primers 1+3). M: marker. *
To whom
correspondence
should be addressed
