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《Acta Academiae Medicine Militaris Tertiae》 2002-01
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Cloning and in vitro expression of human fast-twitch skeletal muscle troponin I gene

XIONG Guang wu, WANG Shi lang, YANG Li, WEI Yu quan, TIAN Ling, LEI Song, KAN Bing, MAO Yong qiu, WANG Yu, YANG Pei, FANG Fang (Center of Biotherapy of Cancer, Huaxi Hospital, Sichuan University, Chengdu 610041, China)  
Objective To construct a recombinant eukaryotic expression plasmid encoding human fast twitch skeletal muscle troponin I gene (TnI fast) for anti angiogenesis in tumor gene therapy. Methods The TnI fast cDNA was amplified with standard PCR and RT PCR by using the human adult normal skeletal muscle cDNA and total RNA as templates, respectively, and then cloned into vector pUC18. After being sequenced in both directions, the cDNA was cloned into the eukaryotic expression vector pSecTag2 B. The obtained recombinant plasmid was transferred to COS 1 cells with a lipofectamine method. The expression of TnI fast in eukaryotic cells in vitro was examined with ELISA. Results DNA sequencing revealed that the PCR and RT PCR amplified TnI fast cDNA was identical to our objective. The recombinant TnI fast/pSecTag2 B plasmid was constructed successfully, and the secreting expression of TnI fast gene was detected in the transfected COS 1 cells. Conclusion Our results indicate that the recombinant TnI fast/pSecTag2 B plasmid can be used for animal study of anti angiogenesis in tumor gene therapy.
【CateGory Index】: R73-3
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