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中国生物工程杂志

China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (12): 46-50    DOI:
    
Cloning and Expression of ompW Location in Outer Membrane of Escherichia coli
ZOU Hai-jie, WU Xian-bin, PAN Jian-yi, ZHAO Fu-kun
School of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China
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Abstract  

Objective:To achieve the mutant stains of Escherichia coli K12 which up-expressed OmpW located in outer membrane by vector pLLP-OmpA. Methods:The complete sequence of ompW from E. coli K12 was cloned by PCR. The cloned DNA fragments were linked with the plasmid to construct the vector pLLP-OmpA-ompW. The recombinant vector was then transferred into E. coli K12 to obtain the engineering strain. Western bolt analysis with primary antibody against OmpW was used to verify whether the highly expressed OmpW of the strain located in outer membrane or not. The polyclonal antibody was prepared by immunizing mice with OmpW protein extracted from strain of pET-28a-ompW. Results: The recombinant expression vector of OmpW was successfully constructed and the high expression strains were also obtained after transformed into the host. The strains which the high-expression of OmpW located in the outer membrane were further confirmed by Western blot analysis. Conclusion: The strain of high-expression proteins location in outer membrane was achieved for the first time. The result will contribute to investigation the function of OmpW.



Key wordsEscherichia coli K12      OmpW      Recombination plasmid      Antiserum preparation     
Received: 03 August 2011      Published: 25 December 2011
ZTFLH:  Q786  
Cite this article:

ZOU Hai-jie, WU Xian-bin, PAN Jian-yi, ZHAO Fu-kun. Cloning and Expression of ompW Location in Outer Membrane of Escherichia coli. China Biotechnology, 2011, 31(12): 46-50.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I12/46


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