©C3 1990 Oxford University Press

6432 Nucleic Acids Research, Vol. 18, No. 21

Nucleotide and deduced amino acid lactate dehydrogenase-A

sequences

of chicken

Y.Hirotal, A.Katsumata and T.Takeya Institute for Chemical Research, Kyoto University, Uji, Kyoto and 'Meiji Institute of Heafth Science, Odawara, Kanagawa, Japan Submitted September 26, 1990

EMBL accession no. X53828

We isolated and sequenced a cDNA clone carrying the lactate dehydrogenase A chain (LDH-A) from a chicken embryo fibroblasts cDNA library by differential hybridization; the expression of the LDH-A was increased eight fold in CEF transformed by Rous sarcoma virus. The cDNA clone is 1575 bp long; it consists of a 59 bp 5' non-coding region, a 996 bp coding region, and a 520 bp 3' non-coding region. The deduced amino acid sequence is identical to the sequence determined by amino acid sequencing (1) except at two residues of Leu-63 and Ile-78 instead of Met-63 and Thr-78, respectively, To investigate the origins of these changes, we further analyzed the nucleotide sequences of the corresponding regions on the genomic clones and found that the genomic sequences of amino acid position 63 and 78 were ACG (Thr) and ACA (Thr), respectively. It thus seems likely that misincorporations occurred at both positions during the cloning process and that the still remaining difference at position 63 (Leu-63 and Thr-63) might be due to other reasons, such as the difference of strain used.

-59

GA ATT CCG CCC CGC TAC GAC CGC AGC CGC GCT AGA AGG GGA AGA CGC CGG CAG TAC ACC

I I

ATG TCT CTC AAG GAT CAT CTC ATC CAC AAT GTC CAC AAA GAG GAG CAC GCT CAT GCC CAC Met Ser Lou Lys Asp ilis Leu Ile His Asn Val Ills Lys Glu Glu His Ala 11is Ala [is

61 21

AAC AAG ATC AGC GTG GTT GGT GTG GGT GCA GTT GGA ATG GCT TGT GCC ATC AGC ATC CTG Asn Lys Ile Ser Val Val Gly Val Gly Ala Val Gly Met Ala Cys Ala Ile Ser Ile Leu

121 41

ATG AAG GAC TTG GCA GAT GAA CTT ACC CTT GTT GAT GTT GTG GAG GAC AAG CTG AAA GGA Met Lys Asp Leu Ala Asp Glu Leu Thr Leu Val Asp Val Val Glu Asp Lys Leu Lys Gly

181 61 241 81

GAA ATG CTT GAT CTC CAG CAT GGC AGC CTC TTC CTC AAA ACA CCA AAG ATT ATA TCT GGC Glu Met Leu Asp Leu Gin His Gly Ser Leu Phe Leu Lys Thr Pro Lys Ile Ile Ser Gly AAA GAT TAC AGC GTG ACT GCA CAC TCC AAG CTG GTC ATT GTC ACT GCT GGT GCC CGT CAG Lys Asp Tyr Ser Val Thr Ala His Ser Lys Leu Val Ile Val Thr Ala Gly Ala Arg Gin

301 101

CAA GAA GGA GAA AGC CGT CTT AAC TTG GTC CAA CGC AAT GTC AAT ATC TTC AAA TTC ATC Gln Glu Gly Glu Ser Arg Leu Asn Leu Val Gin Arg Asn Vol Asn I le Phe Lys Phe Ile

361 121

ATT CCC AAT GTT GTG AAA TAC AGT CCT GAC TGC AAG CTG CTG ATT GTC TCA AAC CCA GTG Ile Pro Asn Val Val Lys Tyr Ser Pro Asp Cys Lys Leu Leu I le Val Ser Asn Pro Val

781 261

GAA ACT ATC ATG AAG AAC TTA AGA AGA GTG CAC CCA ATC TCT ACA GCT GTT AAG CGC ATG Glu Thr Ile Met Lys Asn Leu Arg Arg Val H1is Pro Ile Ser Thr Ala Val Lys GlyI Met

REFERENCES

841 281 901 301

CAC GGA ATA AAA GAT GAT GTC TTC CTA AGT GTT CCT TGT GTA CTG GGC AGC AGT GGC ATC llis Gly Ile Lis Asp Asp Val Phe Leu Ser Val Pro Cys Val Lev Gly Ser Ser GluI le ACT GAT GTA GTG AAG ATG ATC CTA AAA CCT GAT GAA GAG GAG AAA ATA AAG AAG AGC GCA Thr Asp Val Val Lis Met Ile Leu Lis Pro Asp Glu Glu Glu Lis Ile Lys Lys Ser Ala

961 321

GAT ACA CTC TGG GGA ATT CAG AAA GAA CTA CAG TTT TAA AGT TGT CCA AGC AAT TCA CTG Asp Thr Leu Trp Gly le Gin Lis Glu Leu Gin Phe **0

1. Troff,H.J., Becker,D. and Schwarzwalder,J. (1977) In Sund,H. (ed.), Pyridine Nucleotide Dependent Dehydrogenase. Walter de Gruyter, Berlin, pp. 31-42.

421 141

GAT ATT TTG ACC TAT GTG GCC TGG AAG ATC AGT GGC TTT CCT AAA CAC CGT GTT ATC GGT Asp lie Leu Thr Tyr Val Ala Trp Lys Ile Ser Gly Phe Pro Lys His Arg Val Ile ily

481 161

AGT GGC TGC AAT CTG GAC TCA GCC CGT TTC CGC CAC CTC ATG GGA GAA AGA CTG GGC ATC Ser Glu Cis Asn Leu Asp Ser Ala Arg Phe Arg His Leu Met Gly Glu Arg Leu GluI le

541 181

CAT CCT CTG AGC TGC CAT GGG TGG ATT GTT GGA GAG CAT GGA GAC TCC AGT GTG CCT GTC llis Pro Lea Ser Cis 1lis Glu Trp I le Val Glu Glu His Gly Asp Ser Ser Val Pro Val

601 201

Trp Ser Gly Val Asn Val Ala Gly Val Ser Leu Lys Ala Leu 11is Pro Asp Met Gly Thr

661 221

GAT GCA GAC AAG GAG CAC TGG AAG GAG GTT CAC AAG CAG GTG GTG GAC AGT GCC TAT GAG Asp Ala Asp Lys Glu His Trp Lys Glu Val His Lys Gln Val Val Asp Ser Ala Tyr Glu

721 241

GTC ATC AAA CTG AAG GGT TAC ACA TCA TGG GCT ATT GGC CTT TCT GTG GCA GAT CTA GCT Val Ile Lys Leu Lys Gly Tyr Thr Ser Trp Ala le Gly Leu Ser Val Ala Asp Leu Ala

TGG AGC GGA GTG AAT GTT GCT GGT GTC TCC CTG AAG GCT CTT CAT CCA GAC ATG GGA ACT

1021

CAG TTG AAC GGT TTA GTG GTT TGT CTT AAT GTT GCA CTT TCC AAG TAG GTC AAA TCC TTT

1001

GGT GTA TGT ATC TCA ACT AGA AAA CAA ACT AAA ACC TGA ACA AAC GAG TTT CCT AAA GGT

1141

GGC CAT TAG GAA ACT ATT CTA G00

1201

AGC TCC AGC CAC CTA GTA TTG TAC AGT GAA GTG GTG TGA CAT CAA TCC AGC ATT GA0

1261

AGC ACT GCC TTT CCC TCA CCT TGT TTA AAC ATT AGC CCA GAA CAA TAA CTA ACC AGC TAA

1321

CTA ACA CAA CGT GAA ACC AAA GAC TAT TTT CTT GCA GTC ACA GGA TTG TCA AAC TAT TTG

TTT TCC CTG TTC TCA GAC ACT TGT ATT CTT AGC CAG

AGC

1381

ACA AAC CTT CTC TGC COG CTG AAT AGC TTA AAT ATT TCA CTG TTG GTA GGT CCA AAT OTT

1441

TTC CTA ACA CTT CTT TGG AAC TTG TGT ATA CCT TTT TTT TAG AGG AAC TTA TTT TTG TGT

1501

ACT GTA TCC AGA ATT C