[1]贺淹才,刘爱花,柴思捷,等.改进RBB染色法初步筛选几丁质酶基因chiA的表达[J].华侨大学学报(自然科学版),2008,29(4):563-566.[doi:10.11830/ISSN.1000-5013.2008.04.0563]
　HE Yan-cai,LIU Ai-hua,CHAI Si-jie,et al.Improved RBB Dyeing Method Screen Bacteria of Expression of Chintinase ChiA[J].Journal of Huaqiao University(Natural Science),2008,29(4):563-566.[doi:10.11830/ISSN.1000-5013.2008.04.0563]
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改进RBB染色法初步筛选几丁质酶基因chiA的表达()
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《华侨大学学报（自然科学版）》[ISSN:1000-5013/CN:35-1079/N]

卷:
第29卷
期数:
2008年第4期
页码:
563-566
栏目:
出版日期:
2008-10-20

文章信息/Info

Title:
Improved RBB Dyeing Method Screen Bacteria of Expression of Chintinase ChiA
文章编号:
1000-5013(2008)04-0563-04
作者:
贺淹才; 刘爱花; 柴思捷; 刘治江
华侨大学材料科学与工程学院
Author(s):
HE Yan-cai;  LIU Ai-hua;  CHAI Si-jie;  LIU Zhi-jiang
College of Material Science and Engineering, Huaqiao University, Quanzhou 362021, China
关键词:
RBB底物染色;  几丁质酶基因chiA;  粘质沙雷氏菌;  表达
Keywords:
remazol brilliant blue;  chitinase gene chiA;  Serratia marcescens;  expression
分类号:
Q78
DOI:
10.11830/ISSN.1000-5013.2008.04.0563
文献标志码:
A
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.
Abstract:
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-chiA was transferred into E.coli DH5α,the gene was identified,it expressed in BL21(DE3) pLysS,and primary screening was done.

参考文献/References:

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[3] GOMEZ R M, ROJAS A L I. Colloidal chitin stained with Remazol Brilliant Blue RR, a useful substrate to select chitinolytic micro-organisms and to evaluate chitinases [J]. Journal of Microbiological Methods, 2004(2):213-219.doi:10.1016/j.mimet.2003.10.011.
[4] SAMBROOK J, RUSSELL D W. Molecular cloning:A laboratory mamml [M]. 北京:科学出版社, 2002.
[5] 贺淹才, 刘爱花, 张荣奎. 嗜麦芽窄单胞菌产生的几丁质酶的特性 [J]. 华侨大学学报(自然科学版), 2008(2):245-249.
[6] ROBBINS P W, ALBRIGHT C, BENFIELD B. Cloning and expression of a Streptomyces plicatus chitinase(chiti-nase-63)in Escherichia coli [J]. Biochemistry, 1988(3):443-447.

备注/Memo

备注/Memo:
福建省自然科学基金资助项目(C04010011)
更新日期/Last Update: 2014-03-23