摘要
以多聚酶链反应技术扩增获得了杜氏利什曼原虫（Ｌ．ｄｏｎｏｖａｎｉ）热休克蛋白７０（ＨＳＰ７０）全长基因序列，构建ＤＮＡ疫苗表达载体ｐＶＲ１０２０－ＨＳＰ７０。以脂质体法体外转染小鼠成纤维细胞系ＳＶＴ２，应用Ｗｅｓｔｅｒｎｂｌｏｔ法证实构建的表达载体具有表达ＨＳＰ７０的能力。以１０μｇ的ＤＮＡ疫苗表达载体质粒经肌肉、皮下注射进行免疫，２周后即可出现抗－ＨＳＰ７０的抗体应答。建立了ＤＮＡ疫苗免疫应答的模型。
Heat shock protein 70 (HSP70) DNA of Leishmania donovani was amplified by polymerase chain reaction (PCR) and was subcloned into DNA vaccine vector pVR1020 at BamHI site.The expression of HSP70 was confirmed by in vitro transfection of murine fibroblast cell line SVT2 and Western blot hybridization.Antibody response against HSP70 was noticed in the murine serum at 2 weeks after immunization with 10ug of vector plasmid DNA.A mouse model of DNA immunization was established.
出处
《寄生虫与医学昆虫学报》
CAS
1998年第2期84-87,共4页
Acta Parasitologica et Medica Entomologica Sinica
关键词
杜氏利什曼原虫
热休克蛋白
DNA疫苗
转染
PCR
Leishmania donovani\ \ Heat shock protein 70\ \ DNA vaccine\ \ Transfection\ \ Polymerase chain reaction
