Takeuchi H, Shibano Y, Morihara K, Fukushima J, Inami S, Keil B, Gilles A M, Kawamoto S, Okuda K
Department of Bacteriology, Yokohama City University School of Medicine, Japan.
Biochem J. 1992 Feb 1;281 ( Pt 3)(Pt 3):703-8. doi: 10.1042/bj2810703.
The DNA encoding the collagenase of Vibrio alginolyticus was cloned, and its complete nucleotide sequence was determined. When the cloned gene was ligated to pUC18, the Escherichia coli expression vector, bacteria carrying the gene exhibited both collagenase antigen and collagenase activity. The open reading frame from the ATG initiation codon was 2442 bp in length for the collagenase structural gene. The amino acid sequence, deduced from the nucleotide sequence, revealed that the mature collagenase consists of 739 amino acids with an Mr of 81875. The amino acid sequences of 20 polypeptide fragments were completely identical with the deduced amino acid sequences of the collagenase gene. The amino acid composition predicted from the DNA sequence was similar to the chemically determined composition of purified collagenase reported previously. The analyses of both the DNA and amino acid sequences of the collagenase gene were rigorously performed, but we could not detect any significant sequence similarity to other collagenases.
克隆了编码溶藻弧菌胶原酶的DNA,并测定了其完整的核苷酸序列。当将克隆的基因连接到大肠杆菌表达载体pUC18上时,携带该基因的细菌同时表现出胶原酶抗原和胶原酶活性。胶原酶结构基因从ATG起始密码子开始的开放阅读框长度为2442 bp。从核苷酸序列推导的氨基酸序列显示,成熟的胶原酶由739个氨基酸组成,Mr为81875。20个多肽片段的氨基酸序列与胶原酶基因推导的氨基酸序列完全相同。从DNA序列预测的氨基酸组成与先前报道的纯化胶原酶的化学测定组成相似。对胶原酶基因的DNA和氨基酸序列都进行了严格分析,但我们未检测到与其他胶原酶有任何显著的序列相似性。