解藻酸弧菌株ATCC 17749中海藻酸裂解酶基因的克隆与鉴定

Cloning and Identification of Alginate Lyases Genes from Vibrio alginolyticus ATCC 17749

摘要: 以解藻酸弧菌株（Vibrio alginolyticus）ATCC 17749为研究对象，利用生物信息学寻找并预测得到5个可能的海藻酸裂解酶基因algV1、 algV2、 algV3、 algV4和 algV5。通过构建5个以pET 28a(+)为载体的大肠杆菌表达质粒pET28a algV1、pET28a algV2、pET28a algV3、pET28a algV4和pET28a algV5，实现了5个基因的异源表达，并经海藻酸裂解酶定量和定性的活性分析，确定5个基因的编码产物都具有海藻酸裂解酶活性，其中重组的algV1、 algV2和 algV3为胞外酶，algV4和algV5为胞内酶。

Abstract: Finding new sources of alginate lyase gene had a certain scientific research significance and potential application value. In this study, five putative alginate lyase genes were cloned from Vibrio alginolyticus ATCC 17749 according to methods of bioinformatics. Heterologous expressions of five genes in Escherichia coli were achieved by constructing five expression plasmids pET28a algV1，pET28a algV2，pET28a algV3，pET28a algV4，pET28a algV5. Products coded by five genes were verified to own alginate lyase activity by qualitative and quantitative determination. In addition, alginate lyases genes algV1, algV2 and algV3 were extracellular enzyme, and algV4 and algV5 were intracellular enzyme.