Anti-HBV effect of TAT-HBV targeted ribonuclease

查看全文 作　　者：Jin Ding Jun Liu Cai-Fang Xue Wei-Dong Gong Ying-Hui Li Ya Zhao Department of Etiology,Fourth Military Medical University,Xi’an 710032,Shaanxi Province,China

出　　处：《World Journal of Gastroenterology》索引2003年第9卷第7期,共4页

基　　金：National Natural Science Foundation of China,No.30100157;Medical Research Fund of Chinese PLA,No.01MA184;Innovation Project of FMMU.No.CX99005 摘　　要：AIM: To prepare and purify TAT-HBV targeted ribonuclease fusion protein, evaluate its transduction activity and investigate its effect on HBV replication in 2.2.15 cells.METHODS: The prokaryotic expression vector pTAT containing TR gene was used in transforming E. coli BL21(DE3) LysS and TR was expressed with the induction of IPTG. The TAT-TR fusion protein was purified using Ni-NTA-agrose and PD-10 desalting columns, and analyzed by SDSPAGE. Transduction efficiency of TAT-TR was detected with immunofluorescence assay and the concentration of HBeAg in the supernatant of the 2.2.15 cells was determined via solid-phase radioimmunoassay (spRIA). MTT assay was used to detect the cytotoxicity of TAT-TR.RESULTS: The SDS-PAGE showed that the TAT-TR fusion protein was purified successfully, and the purity of TAT-TR was 90 %. The visualization of TAT-TR by immunofiuorescence assay indicated its high efficiency in transducing 2.2.15 cells.RIA result suggests that TAT-TR could inhibit the replication of HBV effectively, it didn′t affect cell growth and had no cytotoxicity.CONCLUSION: TAT-TR possesses a significant anti-HBV activity and the preparation of TAT-TR fusion protein has laid the foundation for the use of TR in the therapeutic trial of HBV infection. 关键词：乙型肝炎病毒核糖核酸酶 TAT融合蛋白蛋白纯化

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Anti-HBV effect of TAT-HBV targeted ribonuclease

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