PCR cloning and sequencing of the beta-amylase cDNA from barley.
Polymerase chain reaction (PCR) amplification of mRNA from developing barley (cultivar Haruna two-rows) endosperm was used to clone and sequence full-length cDNA encoding beta-amylase. The beta-amylase cDNA was 1,775 bp in length. The beta-amylase was deduced to be composed of 535 amino acid residues and its molecular weight was calculated to be 59,610. Kreis et al. reported that the beta-amylase cDNA from barley (cultivar Hiproly) was 1,754 bp in length and coded for a polypeptide of 535 amino acids [Eur. J. Biochem. (1987) 169, 517-525]. A comparison of the beta-amylase sequences from Haruna two-rows and Hiproly barleys revealed nine differences in the nucleotide sequence which resulted in three changes in the amino acid residues and 21 additional nucleotides at its 3'-end in the cultivar Haruna two-rows. The three changes were as follows: Ala-233, Ser-347, Met-527 (Haruna two-rows) and Val-233, Met-347, Ile-527 (Hiproly). Lundgard and Svensson pointed out that 23 amino acid residues of the peptide fragment derived from the COOH-terminal region of barley (cultivar Gula) beta-amylase were in agreement with the deduced amino acid sequence reported by Kreis et al., with the exception of a single position (Met-527 compared to Ile) [Carlsberg Res. Commun. (1986) 51, 487-491]. Our findings described above showed Met-527 is reasonable. In the cases of beta-amylases from soybean and sweet potato, the positions that corresponded to those at 233 and 347 in the amino acid sequence of beta-amylase from barley were Ala and Ser, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
Yoshigi
N
N
Brewing Research Laboratories, Sapporo Breweries Ltd., Shizuoka.
Okada
Y
Y
Sahara
H
H
Koshino
S
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D001467:Hordeum / Q000201:enzymology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D001614:beta-Amylase / Q000737:chemistry* / Q000235:genetics*
D018076:DNA, Complementary; D001614:beta-Amylase