| Project/Area Number |
06680019
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
家政学
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| Research Institution | Nara Women's University |
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Principal Investigator |
YOKOIGAWA Kumio Nara Women's Univ, Faculty of Human Life and Environment, Associate Prof., 生活環境学部, 助教授 (60230637)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAI Hiroyasu Nara Women's Univ, Faculty of Human Life and Environment, Prof., 生活環境学部, 教授 (80026525)
遠藤 金次 聖母女学院短期大学, 生活科学科, 教授 (20031643)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1994: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Alanine racemase / Psychrotroph / Food Hygiene / ポリメラーゼチェインリアクション / 細菌検査 |
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| Research Abstract |
Amplification of a gene fragment of a typical prokaryotic enzyme, alanine racemase, from various bacteria was examined by polymerase chain reaction (PCR) with several degenerate oligonucleotide primers. The primers were designed based upon the conserved sequences of amino acids flanking the Lys 41 and His 168 residues of the enzymes from Bacillus subtilis, Bacillus stearothermophilus, and Salmonella typhimurium. PCR with a pair of primers designed in due consideration of the codon usage in these alanine racemase genes was found to produce a specifically amplified product with a size of approximately 390-base pair from genomic DNAs of Bacillus psychrophilus, Enterobacter cloacae, Bacillus psychrosaccharolyticus, and Escherichia coli. These four amplified products were confirmed as the gene fragments of alanine racemases by DNA sequencing ; the conserved amino acid residues present in known alanine racemases were also found in the four translated sequences of the PCR products. About 390-base pair DNA fragments were also amplified from B.subtilis, B.stearothermophilus, Serratia marcescens, and Pseudomonas fluorescens, but not from a bakers' yeast and Aspergillus niger. The PCR method was applicable to the detection of bacteria in cow's milk : single bacterial cell (Bacillus psychrosaccharolyticus) in cow's milk was detected within 4h.
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