4250 Nucleic Acids Research, Vol. 18, No. 14

© 1990 Oxford University Press

Nucleotide sequence of cDNA for a-amylase from cotyledons of germinating Vigna mungo seeds Daisuke Yamauchi and Takao Minamikawa Department of Biology, Tokyo Metropolitan University, Fukazawa 2-1-1, Setagaya-ku, Tokyo 158, Japan Submitted June 13, 1990

EMBL accession no. X53049

The cDNA expression library of cotyledons of Vigna mungo seedling (1) was screened with the antibody against a-amylase and a cDNA clone (XAMY1) was isolated. A cDNA clone (XAMY11) which contains the complete coding region for aamylase was isolated from the cDNA library in XgtlO (1) with XAMY1 and sequenced. This cDNA insert includes 1263 bp of

open reading frame, and 13 and 249 bp of 5'-and 3'-untranslated regions, respectively. Amino acid sequence determined using the purified enzyme is underlined. FE ENCE REFER 1. Akasofu,H. et al. (1989) Nucl. Acids Res. 17, 6733. CAAAATCAAAGCT

-1

AATT TT CTCCTTC ATIG9ATTC0TCTCGGCTCAG TCTTCCCCTGCCTTGCTGTTTCAGGGGTTC 90 S F

M

S

D

R

L

S

I

F

C

L

F

I

S

AACTGGGAATCAAGTAAGAAAGG GT'GGTACAACIX:X N

W

E

S

S

K

K

G

G

W

Y

N

S

L

L

L

P

L

AGAACTCTATTCC K

N

S

I

P

F

S

S

P

A

L

L

F

Q

G

F

ACCTTGCAAATGCAGGAATCACTCATGTGTGG D

L

A

N

A

G

I

T

II

V

W

30

180 60

CTCCCTCC2CCCTCTCAATCTGT7rCTCCTGAAGGATATCTTCCAGGAAGGCTATATGATC0l'GATGCATCAAAATAlSGATCCAAGAAT 270 L P P P S V Q

S

S

P

E

G

Y

L

P

G

R

L

Y

D

L

D

A

S

K

Y

G

S

K

N

90

GAACTGAAGTCCCTGATTGCTGCrTTTCATGAGAAGGGTATAAAGTGCCTAGCTGACATAGTGATCAACCACAGAACTGCAGAGAGGAAG E L K S L I A A F

360 120

GATGGGAGAGGAATATATTGTA1'TTTGAAGGTGGTACTCCAGATTCACGCCAAGATTGGGGTCCATCTTTCATTTGCAGAGATGACACT D G R G I Y C I F E G G T P D S R D P

450 150

GCATATTCTGATGGCACTGGAAA'AATGACAGTGGAGAGGGATATGATGCTGCACCTGACATTGATCATCTCAATCCACAGGTGCAAAGA A Y S D G T N

540 180

IH

E

K

G

I

K

C

L

A

D

Q

G

N

D

S

G

E

G

Y

D

A

A

I

W

P

D

V

I

G I

N

S

D

H1

F

R

I

If L N

T

C

P

A

R

Q

E

D

V

R

D

Q

K

T

R

GAGTTGTCAGAATGGATGAACTG;CTCAAAACTGAGATTGGTTTTGATGGATGGAGGTTTGA30TGTGAAGGGCTATGCCCCTAGcATc 630 E L S E W M N W K L

T

E

I

G

F

D

G

W

R

F

D

F

V

K

G

Y

A

P

S

I

210

TC(:AAGATCTATATGGAACAGAC rAAGccAGATm2cAGTAGGTGAGAAATGGGATTCTATCTCATATGGGCAGGAT0GGGAAGCCAAAC 720 S

K

I

Y

M

E

Q

T

K

P

D

F

A

V

G

E

K

W

D

S

I

S

Y

G

Q

D

G

K

P

N

TATAACCAAGATAGTCATCGTGG 3GCACTGGTGAATTGGGTTGAATCTGCAGGTGGAGCTATTACTGCATTTGATTTCACAACAAAAGGA Y

N

Q

D

S

H

R

G

A

L

V

N

W

V

E

S

A

G

G

A

I

T

A

F

D

F

T

T

K

G

240 810 270

ArrCTTCAGGCCTGTGCAAGG.%GAATTGTGGAGGTTGATAGATCCAAATGGCAAGCCTCCTGGAATGATTGGTGTGAAACCAGAAAAT I L Q A A V Q

900 300

GCAGTAACTTTCATTGATAACCATGACACTGGTTCCACACAGAGACTTTGGCCATTCCCTTCTGACAAAGTCATGCAAGGATAT'GCCTAC A V T F I D N If D T F

990 330

ATTCTAACACACCCTGGCACCCC -TAATATTCTACGACCACTTCTlTTATTGGGGCTTGAAGGAGCAGATTGCAAAATTGAGTAGCATA

1080

G

E

L

W

G

I

L

T

II

P

G

T

P

S

I

F

R

S

Y

L

T

D

I

Q

H

D

R

F

P

L

F

N

W

D

G

K

P

W

P

P

G

L

P

S

K

G

D

E

M

K

Q

I

V

I

G

M

A

V

Q

K

K

G

L

P

Y

S

E

A

S

N

Y

I

360

AGGTTGAGGAATGGGATCAATGAGAAAAGCACTGTTAAAATTATGGCATCTGAAGGTGACCTTTATGTTGCAAAGATAGACAACAAGATC R L R N G I N E K S T V K I M A S E G

1170 390

Al'GGTGAAGATAGGGCCAAAGAT-3GACCTTGGAAATCTT'ATTCCCTCAAATCTCCATG'MTCTACGTCTGGCCAAGACTATGCCGTGTGG V K I G P K N 13 L G N L I P S N L 1 V A T S G Q D Y A V

1 26 0 420

D

L

Y

V

A

K

I

N

D

N

K

I

W

G1G35GAATAATTTATTGCAGTAr0CATACAATAAGGAACAC0-TCATGTrrATCATAAAC'lACClwlsl'CATTGGGTATAAGATCATrrAT 1350 E 421

AATTCACCCC

ACCCAATGCA

1440 T144CCCT0AATATATAAACCATAATTCCAAGATCACAAATCATTArGCAAAATTTTCATArr

ACTTGTGTGAGAAATTCGATGAAAAGATGGTATT,GTTATSlTAAATCGATCTCCTCAAGAGTTATTATT

pol yA

1 52 5