Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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A Gene Encoding $\beta$-amylase from Saprolegnia parasitica and Its Expression in Saccharomyces cerevisiae

  • Kim, Hee-Ok (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Park, Jeong-Nam (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Shin, Dong-Jun (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Lee, HwangHee Blaise (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Chun, Soon-Bai (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Bai, Suk (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University)
  • Published : 2001.06.01
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Abstract

The ${\beta}$-Amylase cDNA fragment from the oomcete Saprolegnia parasitica was cloned by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from conserved ${\beta}$-amylase sequences. The 5'and 3'regions of the $\beta$-amylase gene were amplified using the rapid amplification of cDNA ends (rACE) system. It consisted of an open reading frame of 1,350 bp 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 ${\beta}$-amylase gene had a 97% similarity to the ${\beta}$-amylase of Saprolegnia ferax, followed by 41% similarity to those of Arabidopsis thaliana, Hordeum vulgare, and Zea mays. The ${\beta}$-amylase gene was also expressed in Saccharomyces cerevisiae by placing it under the control of the alcohol dehydrogenase gene (ADC1) promoter.

Keywords

References

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