VASP基因RNA干扰慢病毒表达载体的构建与鉴定

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Abstract Objective To construct the lentiviral RNA interference expression vector targeting vasodilator-stimulated phosphoprotein (VASP) gene. Methods The pre-constructed pcDNA6.2-miRVASP was identified by DNA sequencing and the positive clones were screened. The miRVASP fragment was cloned into destination vector pLenti6/V5-DEST by Gateway techology, including BP and LR reaction, RNA interference lentiviral vector pLenti6/V5-miRVASP targeting VASP gene was constructed. Then the 293FT cells were co-transfected with the plasmid lentiviral expression vector pLenti6/V5-miRVASP and lentiviral packaging mix. The virus particles were collected and infected into gastric cancer BGC-823 cells. Results The constructed lentiviral expression vector pLenti6/V5-miRVASP was introduced into E.coli Stbl3 for amplification. DNA sequencing results showed that the constructed vector was exactly as expected. BGC-823 cells were infected with the collected lentiviral stocks carrying the miRVASP expression cassette. The results showed that a large number of cells could express strong green fluorescence, demonstrated that the packaged lentiviral stocks could infect BGC-823 cells. Conclusion The RNA interference lentiviral vector targeting VASP gene is successfully constructed and packaged as the lentiviral stocks to successfully infect BGC-823 cells, which lay the foundation for the further study of the role of VASP in gastric cancer metastasis in vivo.

Key words： VASP RNAi Lentiviral vector

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	WANG Jing1 WEI Lei2 QIU Kun1

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WANG Jing1 WEI Lei2 QIU Kun1. Construction of lentivirus-mediated RNAi vector targeting vasodilator-stimulated phosphoprotein[J]. 中国医药导报, 2018, 15(25): 9-12.

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http://www.yiyaodaobao.com.cn/EN/ OR http://www.yiyaodaobao.com.cn/EN/Y2018/V15/I25/9

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