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Bacillus sp . E1 의 cyclodextrin 생산효소 유전자 분리 및 구명
용정식,최진남,박성순,박천석,박관화,최양도 ( Jeongsik Yong,Jin Nam Choi,Sung Soon Park,Cheon Seok Park,Kwan Hwa Park,Yang Do Choi ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.6
To isolate a gene for cyclodextrin glycosyltransferase (CGTase) from alkalophilic Bacillus sp. El, polymerase chain reaction (PCR) amplification was carried out. Direct molecular cloning of 1.2 kbp fragment and partial nucleotide sequence analysis of the PCR amplified clone, pH12, showed close homology with CGTases from Bacillus species. To investigate the genomic structure of the gene, Southern blot analysis of genomic DNA was carried out with the clone pH12 as a molecular probe. It showed that 5.3 kbp XbaI fragment was hybridized with the probe pH12. To isolate a genomic clone, genomic DNA library was constructed and a genomic clone for CGTase, pCGTEl, was isolated. Nucleotide sequence analysis of the clone pCGTEl revealed that BCGTEl contained 2,109 bp open reading frame encoding a polypeptide of 703 amino acids and showed over 94.3% amino acid sequence homology with CGTase of β-cyclodextrin producer, Bacillus sp. KC201.