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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (10): 83-87    DOI:
    
Knockout of waaL Gene of Escherichia coli O157:H7 by Red Recombination System
YANG Gong-jin, MA Zhong-rui, XIN Guo, CHEN Min
National Glycoengineering Research Center of The State Key Laboratory of Microbial Technology, College of Life Science, Shandong University, Jinan 250100, China
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Abstract  

Objective: To construct a waaL knockout mutant of Escherichia coli O157:H7. Methods: Using plasmid pKD4 as a template,the kanamycin-resistant gene flanked by homologues of waaL gene was amplified by PCR. The PCR products were electro-transferred into the O157:H7,with the help of the Red recombinant system,WaaL gene were replaced by the kanamycin-resistant gene. Then the kanamycin-resistant gene was eliminated by FLP-promoted recombination system. Result: waaL gene of the E. coli O157:H7 was completly eliminated and the kanamycin-resistant gene was also eliminated.

Key words： O157:H7      λ-Red      recombination system      WaaL gene     
Received: 12 May 2011      Published: 25 October 2011
ZTFLH:  Q789  
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Cite this article:

YANG Gong-jin, MA Zhong-rui, XIN Guo, CHEN Min. Knockout of waaL Gene of Escherichia coli O157:H7 by Red Recombination System. China Biotechnology, 2011, 31(10): 83-87.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I10/83


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