MOLECULAR-CLONING AND NUCLEOTIDE-SEQUENCE OF THE LIPASE GENE FROM PSEUDOMONAS-FRAGI
MOLECULAR-CLONING AND NUCLEOTIDE-SEQUENCE OF THE LIPASE GENE FROM PSEUDOMONAS-FRAGI
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DOI：
10.1016/s0006-291x(86)80352-7
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发表时间：
1986-11-26
影响因子：
3.1
通讯作者：
TAKAGI, Y
中科院分区：
文献类型：
--
作者：
KUGIMIYA, W;OTANI, Y;TAKAGI, Y
通过将Sau3A产生的DNA片段插入pUC 9的BamHI位点，将编码脆弱假单胞菌脂肪酶的基因克隆到大肠杆菌JM 83中。对分离的质粒pKKO进行限制性作图，并通过亚克隆确定脂肪酶基因在2.0 kb插入片段上的位置。DNA序列分析表明，开放阅读框架包括405个核苷酸，并给出了一个135个氨基酸的前蛋白，预测的Mr为14643。通过比较推定的脂肪酶的氨基酸序列与猪胰，大鼠舌和葡萄球菌的脂肪酶的反应性丝氨酸周围的氨基酸序列被发现是常见的脂肪酶的类型，已报告。
The gene coding for the lipase of Pseudomonas fragi was cloned into Escherichia coli JM83 by inserting Sau3A-generated DNA fragments in the BamHI site of pUC9. The plasmid isolated, pKKO, was restriction mapped and the position of the lipase gene on the 2.0 kb insert was pinpointed by subcloning. DNA sequencing revealed that the open reading frame comprises 405 nucleotides and gives a preprotein of 135 amino acids with a predicted Mr of 14643. By comparing the putative lipase amino acid sequence with porcine pancreatic, rat lingual and Staphylococcus hyicus lipases the amino acid sequence around the reactive serine was found to be common among the types of lipase which have been reported.