【摘要】 以猪瘟病毒石门株RNA序列为模板,应用RT-PCR方法,克隆得到猪瘟病毒囊膜蛋白E0-E2cDNA全序列,并将其与真核表达质粒pcDNA4.0定向连接,构建了重组质粒pcDNA4.0-E。经提纯后,给试验组小鼠后肢胫前肌注射pcDNA4.0-E,每只100μg,15 d后再注射1次,同时设空白对照组。于二免后第10、203、0 d分别扑杀小鼠,进行免疫小鼠脾和外周血淋巴细胞的转化增殖试验,并用间接ELISA法检测血清抗体水平。结果显示,与空白对照组相比,所构建的重组质粒能够极显著增强小鼠脾细胞和外周血淋巴细胞的增殖(P<0.01),血清抗体水平从二免后的第10 d开始逐渐升高,且极显著高于对照组(P<0.01)。说明该重组质粒能够诱导小鼠产生特异性免疫反应。更多还原

【Abstract】 The full-length cDNA encoding envelope protein of classical swine fever virus(CSFV)was amplified by RT-PCR from the RNA sequence of CSFV Shimen strain,and the amplified product was cloned into eukaryotic expression plasmid pcDNA4.0 to construct recombinant plasmid pcDNA4.0-E.Then,9 Kunming mice were inoculated with the purified pcDNA4.0-E in hind legs by intramuscular injection(100 μg per mouse),and the second injection was carried out after 15 days.The indirect ELISA assay showed that the specific antibody levels in sera isolated from peripheral blood of the immunized-mice,which were killed on 10th,20th and 30th day post-2nd-immunization respectively,were gradually increased from 10th day and were significantly higher than those of the non-treatment control group(P<0.01).MTT assay showed that the pcDNA4.0-E expressed in the immunized-mice could obviously enhance lymphocyte proliferation in the spleen and the peripheral blood of the immunized mice compared with those of the control group(P<0.01),respectively.These results showed that the expressed envelope protein could induce specific immunoreaction.更多还原