HCV核心蛋白在昆虫细胞中的表达及其免疫学特性的初步评价

通过逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）从丙肝患者的血清中分离出编码完整ＨＣＶ核心蛋白（Ｃ区）的ｃＤＮＡ片段，并将其克隆到杆状病毒转移质粒中。重组转移质粒ＤＮＡ与线性的杆状病毒ＤＮＡ共转染Ｓｆ９昆虫细胞，经蚀斑筛选获得了带编码全部核心蛋白基因的重组杆状病毒。重组病毒感染细胞后表达ＨＣＶ核心蛋白，其分子量的为２０ｋＤ。免疫印染和酶联免疫实验表明，此重组蛋白能被人ＨＣＶ阳性血清所识别。动物实验表明此重组蛋白能诱导小鼠产生特异性抗体。

Expression of HCV Core Protein in Insect Cells and Evaluation of Its Immunological properties

he full-length cDNA coding HCV core protein was isolated from human HCV positiveserum by RT-PCR and inserted into baculovirus transfer vector.Sf9 insect cells were co-transfect-ed with the recombinant transfer vector plasmid and wild type baculovirus DNA.The recombi-nant baculoviruses containing gene of HCV core protein were obtained by plaque screening.Therecombinant HCV core proteins(20 kD)were expressed by Sf9 cells infected with recombinantviruses.Western blot and EIA revealed that the recombinant proteirls can be recognized by humanHCV positive sera,and the recombinant protein ean elicited specific anti-HCV antibody in ani-mals.