頁籤選單縮合
題名 | Cloning, Sequencing, and Expression of the Creatinase Gene from Pseudomonas putida RS65 in Escherichia coli= |
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作者 | Liu,Wen-hsiung; Tang,Tsung-yin; Wen,Chi-jen; Lin,Yu-cheng; |
期刊 | Food Science and Agricultural Chemistry |
出版日期 | 19990400 |
卷期 | 1:2 1999.04[民88.04] |
頁次 | 頁115-121 |
分類號 | 361.19 |
語文 | eng |
關鍵詞 | Creatinase; Creatine amidinohydrolase; Pseudomonas putida; Cloning; DNA sequencing; |
英文摘要 | A gene encoding creatinase from P. putida RS65 was cloned, sequenced and expressed in E. coli. One plasmid containing a 7.0-kb Hind Ⅲ insert was selected by its ability to express creatinase activity. The DNA insert of this plasmid was mapped by restriction enzymes. The creatinase gene was subcloned in a 2.7-kb Sal I-Eco R I fragment by deleting the adjacent restriction fragments. The nucleotide sequence of the creatinase gene was determined. The gene consists of 1218 base pairs and encodes a protein of 406 amino acids, with a calculated molecular weight of 46,309. The constitutive creatinase productivity of E. coli DH5 α transformant (pCLN742) cultured in a Luria-Bertani medium was about 35-fold higher than that of P. putida RS65 cultured in a creatine-induced medium. |
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