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KMID : 0545120010110030529
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Journal of Microbiology and Biotechnology
2001 Volume.11 No. 3 p.529 ~ p.533
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A Gene Encoding β-Amylase from Saprolegnia parasitica and Its Expression in Saccharomyces cerevisiae
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Kim, Hee Ok
Park, Jeong Nam/Shin, Dong Jun/Lee, Hwanghee Blaise/Chun, Soon Bai/Bai, Suk
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Abstract
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The P-Amylase cDNA fragment from the oomycete Saprolegnia parasitica was cloned by reverse transcriptionpolymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from conserved β-amylase sequences. The 5´ and 3´ regions of the β-amylase gene were amplified using the rapid amplification of cDNA ends (RACE) system. It consisted of an open reading frame of 1,350bp for a protein of 450 amino acids. Comparison between the genomic and cDNA sequences revealed that the intron was not present in the coding region. The deduced amino acid sequence of the β-amylase gene had a 97% similarity to the β-amylase of Saprolegnia ferax, followed by 41% similarity to those of Arabidopsis thaliana, Hordeum vulgare, and Zea mays. The β-amylase gene was also expressed in Saccharomyces cerevisiae by placing it under the control of the alcohol dehydrogenase gene (ADCl) promoter.
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KEYWORD
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