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| Construction of Brucella Deletion Mutant by Using T Cloning Vector |
| BAI Yao-xia1,2,YANG Yi2,WANG Yu-fei2,WANG Tong-kun2,YU Shuang2,CHEN Yan-fen2,FU Si-mei2,HUANG Liu-yu2,TIAN Jin-hong1,CHENG Ze-liang2 |
1.College of Pharmaceutical Science, Southwest University, Chongqing 400716,China
2.Institute of Disease Control and Prevention, Academy of Military Medical Science, Beijing 100071,China |
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Abstract Control and Prevention, Academy of Military Medical Science, Beijing, 100071） Abstract: Construction of mutant strain is an essential method in pathogenesis researches. In the present study, the Brucella deletion mutant was constructed by using T cloning vector. At first, the homology arms of upstream and downstream of the target gene were fused with kanamycin resistance gene by fusion PCR. Then, the mutant cassette was ligated with T vector, resulted in the mutant plasmid. And the mutant plasmid was transformed into recombination competent cells and recombinants were selected by kanamycin resistance. As the results showed, the mutant construction was convenient and the mutant could be generated by only one round of selection with high efficiency. Therefore, this recombinational cloning strategy provides us a new method to construct the Brucella mutant and would accelerate gene function analysis of Brucella.
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Received: 04 May 2010
Published: 25 August 2010
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Cite this article:
BAI Yao-Xia, YANG Yi, WANG Yu-Fei, WANG Tong-Kun, XU Shuang, CHEN Yan-Fen, FU Sai-Mei, HUANG Liu-Yu, TIAN Jin-Gong, CHEN Ze-Liang. Construction of Brucella Deletion Mutant by Using T Cloning Vector. China Biotechnology, 2010, 30(09): 62-67.
URL:
https://manu60.magtech.com.cn/biotech/Q785 OR https://manu60.magtech.com.cn/biotech/Y2010/V30/I09/62
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