负链RNA病毒反向遗传通用载体的构建与鉴定

doi:10.3969/j.issn.1004-616x.2010.03.004

Carcinogenesis, Teratogenesis & Mutagenesis ›› 2010, Vol. 22 ›› Issue (3): 175-178.doi: 10.3969/j.issn.1004-616x.2010.03.004

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Construction of universal reverse genetic vector for minus RNA viruses

WANG Ge-fei;ZHANG Chi;ZENG Xiang-xing;ZHANG Heng;CHEN You-ying;LI Wei-zhong;LI Kang-sheng

Department of Microbiology and Immunology, Key Immunopathology Lab of Guangdong Province,Shantou University Medical College,Shantou 515041, Guangdong, China

Received:2009-07-09 Revised:2009-12-28 Online:2010-05-30 Published:2010-05-30
Contact: LI Kang-sheng

Abstract

Abstract: OBJECTIVE: To construct a system for reverse genetics of minus RNA viruses, the universal reverse genetic vector was constructed. METHODS: Using pcDNA3 as plasmid backbone, non-essential Kanamycin resistant expression cassettes and CMV promoter upstream region were removed. Then the fragment including reverse pol I promoter and multiple clone site were synthesized by PCR and inserted into the reconstructed pcDNA3. The modified pcDNA3 and universal reverse genetic vector were identified by restriction enzyme. RESULTS: Sequencing showed that the universal reverse genetic vector which had CMV and pol I bidirectional promoter was constructed successfully. CONCLUSION: This research established reverse genetics system of minus RNA viruses and provided research basis for further development of minus RNA viruses.

Key words: reverse genetics, RNA virus, vector

CLC Number:

R373.1，Q939.4

WANG Ge-fei, ZHANG Chi, ZENG Xiang-xing, ZHANG Heng, CHEN You-ying, LI Wei-zhong, LI Kang-sheng. Construction of universal reverse genetic vector for minus RNA viruses[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2010, 22(3): 175-178.

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Construction of universal reverse genetic vector for minus RNA viruses

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