A gene encoding Achlya bisexualis β-amylase and its expression in Saccharomyces cerevisiae

Part of a beta -amylase genomic DNA sequence from the oomycete, Achlya bisexualis was cloned by polymerase chain reaction (PCR) using degenerate oligonucleotide primers derived from the conserved regions of other known beta -amylase sequences. The 5'- and 3'-regions of the beta -amylase ge...

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Bibliographic Details
Published in:Biotechnology letters 2001-07, Vol.23 (14), p.1101-1107
Main Authors: KIM, Hee-Ok, PARK, Jeong-Nam, SHIN, Dong-Jun, LEE, Hwanghee Blaise, CHUN, Soon-Bai, SUK BAI
Format: Article
Language:English
Subjects:
Achlya bisexualis
Biological and medical sciences
Biology of microorganisms of confirmed or potential industrial interest
Biotechnology
DNA sequencing
Fundamental and applied biological sciences. Psychology
Miscellaneous
Mission oriented research
Saccharomyces cerevisiae
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Summary:Part of a beta -amylase genomic DNA sequence from the oomycete, Achlya bisexualis was cloned by polymerase chain reaction (PCR) using degenerate oligonucleotide primers derived from the conserved regions of other known beta -amylase sequences. The 5'- and 3'-regions of the beta -amylase gene were amplified by genome walking method. The Ach. bisexualis beta -amylase gene consisted of a 1338 bp open reading frame, encoding a protein of 446 amino acids with a molecular weight of 49 381 Da, and was not interrupted by any intron. The deduced amino acid sequence of the beta -amylase gene had 67% similarity to the beta -amylase of Saprolegnia ferax, followed by 40% similarity to that of Arabidopsis thaliana. The beta -amylase gene was expressed in Saccharomyces cerevisiae placing it under the control of the alcohol dehydrogenase gene (ADC1) promoter.
ISSN:0141-5492
DOI:10.1023/A:1010503731486