全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

改进RBB染色法初步筛选几丁质酶基因chiA的表达

DOI: 10.11830/ISSN.1000-5013.2008.04.0563

Keywords: RBB底物染色, 几丁质酶基因chiA, 粘质沙雷氏菌, 表达

Full-Text   Cite this paper   Add to My Lib

Abstract:

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.

References

[1]  CHEN H C, CHANG C C, MAU W J. Evaluation of N-acetylchitooligo.saccharides as the main carbon sources for the growth of intestinal bacteria [J]. FEMS Microbiology Letters, 2002(1):53-56.doi:10.1111/j.1574-6968.2002.tb11108.x.
[2]  UWE T B, JOSEF A, HARTMUT H M. Directed evolution of an esterase:Sereaning of enzyme libraries based on pH-indicators and a growth assay [J]. Bioorganic and Medicinal Chemistry Letters, 1999(6):2169-2173.
[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.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133