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| Construction of B.ovis virB Mutant and Analysis the Intracellular Environmental Adaptations |
| GAO Guang-jun1,2, KE Yue-hua1, XU Jie2,3, WANG Li-ping1,2, ZHANG Zhen-fang1,2, LI Zhuo-ling2, GUO Ying-fei2, WANG Lu-lu2, WANG Yu-fei2, XU Xing-ran1, CHEN 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;
3. School of Public Health, Research Center for Genomic Medicine, Jilin University, Changchun 130021, China |
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Abstract Objective: To construct a deletion mutant of virB of B. ovis for functional analysis of Type IV secretion system by using T vector based rapid cloning technique. Methods: The promoter region of virB was deleted by Kanamycin gene through homologous recombination. Transcription of virB genes were verified by RT-PCR. The survival capabilities of the mutant under stress conditions simulating intracellular environments were tested. Results: A deletion mutant of virB was successfully constructed. Expression of the virB genes were inhibited in the mutant. Compared with the wild type strains, the mutant showed altered survival capabilities under stress conditions. Conclusion: A deletion mutant of virB was successfully constructed by using T vector cloning and resistance gene replacement. This provide basis for further functional analysis of type IV secretion system.
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Received: 08 June 2012
Published: 25 November 2012
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Cite this article:
GAO Guang-jun, KE Yue-hua, XU Jie, WANG Li-ping, ZHANG Zhen-fang, LI Zhuo-ling, GUO Ying-fei, WANG Lu-lu, WANG Yu-fei, XU Xing-ran, CHEN Ze-liang. Construction of B.ovis virB Mutant and Analysis the Intracellular Environmental Adaptations. China Biotechnology, 2012, 32(11): 86-91.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2012/V32/I11/86
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