伪狂犬病病毒Ea株糖蛋白gD基因的表达及基因免疫

Expression and Genic Immunization of Glycoprotein gD gene of Pseudorabies Virus Ea Strain

构建了 2个利用人类巨细胞病毒 ( HCMV)的启动子启动表达伪狂犬病病毒 Ea株糖蛋白 g D基因的真核表达质粒 p CIDI和 pc DDI,体外转染 BHK-2 1细胞 ,用间接免疫荧光法检测 ,证实糖蛋白 g D在细胞中得到表达。用表达质粒 p CIDI和 pc DDI作为核酸疫苗免疫 BALB/c小鼠 ,ELISA检测小鼠血清中抗伪狂犬病病毒的抗体 ,结果其滴度为 1∶ 1 2 8～ 1∶ 51 2。初步证实 ,用 g D基因作为核酸疫苗免疫动物 。

Two eukaryotic expression recombinant plasmids,pCIDI and pcDDI containing glycoprotein gD gene of pseudorabies virus Ea strain,were constructed respectively.The expression of foreign gene was promoted by immediate early enhancer/promoter of human cytomegalo virus.Both plasmids were transfected into BHK 21 cell in vitro .Expressed protein was detected by indirect immunofluorescence assay.The recombinant plasmids were also used as genic vaccines to inject BALB/c mice intramuscularly with 100 μg/mouse.The immunization was boosted at 7th week after first injection.Serum samples were collected in 1st,5th,12th week after first injection,respectively.The antibody titers were measured by indirect ELISA to be 1∶128~1∶512.