HCV多表位复合抗原基因表达条件的探索  

Establishment of conditions for HCV multi-epitope antigen gene expression

作　　者：杨蓉[1] 杨婷[1] 李华[1] 龙润乡[1] 董承红[1] 董丽娟[1] 谢忠平[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所,昆明市650118

出　　处：《实用医学杂志》2012年第18期3012-3015,共4页The Journal of Practical Medicine

基　　金：国家863项目(编号:2007AA02Z480);云南省联合支持国家计划项目(编号:2008GA008)

摘　　要：目的:筛选出适合HCV多表位复合抗原基因(PCX)表达的菌株,优化表达条件,并进行目的蛋白的免疫特异性的检测。方法:将含HCV多表位复合抗原基因的重组质粒pWR450-PCX转入大肠杆菌DH5α、TOP10F′、BL21菌株中,在不同培养基中,经不同浓度IPTG诱导不同时间,表达融合蛋白(GZ-PCX),并利用患者阳性血清通过Western Blot检测其免疫特异性。结果:重组质粒在DH5α、TOP10F′菌株中均能表达有一定免疫特异性的目的蛋白,但BL21中不表达;目的蛋白在TB培养基中的表达量明显高于LB培养基。结论:在37℃,IPTG浓度为1mM诱导3h时,目的蛋白的表达量达到峰值;所制备的抗原具有一定的免疫特异性和免疫原性。Objective To screen E.coli strain for HCV muhi-epitope antigen gene （PCX） expression, and to optimize conditions for PCX expression and to detect the immune specificity of the expressed product. Methods The recombinant plasmid, pWR450-PCR, containing HCV multi-epitope antigen gene was transformed into DH5α, TOP10F′ and BL21 strains,respectively, and the fusion protein （GZ-PCX） was induced to express by different concentrations of IPTG for different periods and in different medium. Then the immune specificity of the expressed protein was identified by Western blotting assay with the HCV positive serum. Results The recombinant plasmid, which was transformed into DH5α, TOP10F′ strains, could express the fusion protein with immune specificity, but the recombinant plasmid transformed into BL21 strains failed to express corresponding product. The target protein expression in the TB medium was significantly higher than that in the LB medium. Conclusion The target protein can be expressed efficiently at 3 houra after 1 mM IPTG induction at 37℃. The antigen, prepared has a certain immune specificity and immunogenicity.

关 键 词：丙型肝炎病毒 原核表达 免疫特异性 免疫原性 

分 类 号：R392[医药卫生—免疫学]