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.
出处
《华侨大学学报（自然科学版）》
CAS
北大核心
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)
