改进RBB染色法初步筛选几丁质酶基因chiA的表达  

Improved RBB Dyeing Method Screen Bacteria of Expression of Chintinase ChiA

作　　者：贺淹才[1] 刘爱花[1] 柴思捷[1] 刘治江[1] 

机构地区：[1]华侨大学材料科学与工程学院,福建泉州362021

出　　处：《华侨大学学报（自然科学版）》2008年第4期563-566,共4页Journal of Huaqiao University(Natural Science)

基　　金：福建省自然科学基金资助项目(C04010011)

摘　　要：对RBB(Remazol Brilliant Blue)底物染色方法进行产几丁质酶细菌的酶活性筛选,应用几丁质胶体平板诱导表达的筛选取得好的效果.粘质沙雷氏菌几丁质酶基因chiA在E.coli中的异源表达过程:首先从质粒pMD-chiA(DH5α)中切出含粘质沙雷氏菌几丁质酶基因chiA的片段,将其插入到表达载体pET-22b(+)内,构建成几丁质酶表达质粒pET-chiA.然后,转化进入DH5α感受态细胞内,鉴定正确后将pET-chiA质粒转入BL21(DE3)pLysS中进行表达,并进行初步筛选.Remazol brilliant blue （RBB） dyeing method was improved and used to screen and induce chintinase-producing bacteria in colloidal chitin medium of plate-culture. It got useful results. In order to express Serratia marcescens chitinase gene chiA in E. coli chiA fragment from Serratia marcescens was isolated from the plasmid pMD-chiA（DH5α） and inserted into the expression vector pET-22b（＋）, yielding the expression plasmid pET-chiA. Then the plasmid pET-ehiA was transferred into E. coli DH5α, the gene was identified, it expressed in BL21 （DE3） pLysS, and primary screening was done.

关 键 词：RBB底物染色 几丁质酶基因chiA 粘质沙雷氏菌 表达 

分 类 号：Q-336[生物学] S572[农业科学—烟草工业]