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Establishment of conditions for HCV multi-epitope antigen gene expression
Author(s): 
Pages: 3012-3015
Year: Issue:  18
Journal: The Journal of Practical Medicine

Keyword:  丙型肝炎病毒原核表达免疫特异性免疫原性;
Abstract: 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1 mM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.
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