MS2噬菌体壳蛋白载动的HIV反链基因阻断病毒蛋白质合成  

INHIBITING OF SYNTHESIS OF HIV PROTEIN USING ANTI-SENSE TAT GENE CARRIED BY BACTERIOPHAGE CAPSIDS

作　　者：吴敏[1] 

机构地区：[1]泸州医学院微生物教研室,英国利兹大学

出　　处：《中华微生物学和免疫学杂志》1994年第5期292-295,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金;卫生部青年基金

摘　　要：作者将ＭＳ２噬菌体壳蛋白基因连接于ｐＴＡＣＨ质粒，以Ｅ．ｃｏｌｉＴＧｌ株表达出融合蛋白，用硫酸铵粗提蔗糖梯度离心纯化，通过高浓度冰乙酸使壳蛋白解体，用ＴＭＫ缓冲液中和，与ＴＡＴ反链基因片段混合反应，使之包裹于ＭＳ２壳蛋白。以化学修饰法使此装配的ＭＳ２带上游离巯基，在转铁蛋白的氨基上接入马来酰胺键。藉此二键连成ＭＳ２－ＴＡＴ－ＴＦ复合物。在氯霉素乙酰转移酶试验中，此导向基因治疗剂对ＨＩＶ的ＴＡＴ基因转染的ＨｅＬａ细胞有明显的抑制增植作用，提示本基因治疗剂可应用于预防和治疗ＨＩＶ感染，即ＭＳ２噬菌体壳蛋白作为反链基因载体显出一定前景。S2 coat protein gene was introduced into the pTACH-ACP'which can produce MS2 coat protein in an E.coli.MS2 coat protein expressed by E. coli was purified using saturated ammonium sulfate ,and 15%～45%su-crose gradient centrifugation.MS2 coat protein was dissociated with acetic acid,and changed to TMK buffer(tris-HCl 100mmol/L;MgCl 10mmol/L,KCl 80mmol/L),then mixed with anti-sense TAT oligonucleotide(to the first22 nucleotides of TAT in HIV LTR region)which is joined with MS2 minimum TR sequence,and reacted as fol-lowing:37℃ for 3h,room temperature(20℃)for 3h and 4℃ at least overnight.The encapsulated MS2-TAT wasmdidified to get a thio-group,and meantime transferrin(TF)introduced with a maleimidic group.The two a chemi-cally- modified molecules were then mixed to form conjugates,and confirmed by SDS-PAGE,Western blot,and electron microscope. The results indicated that the package was a specific inhibitor protein synthesis over those controls,and thus may have potential to treat AIDS in animal or in humans.

关 键 词：反链治疗剂 MS2噬菌体 壳蛋白 合成 艾滋病 

分 类 号：R392－33[医药卫生—免疫学]