摘要
The gene fragment coding for amino acids 281 to 395 of the E protein of DENV-2 (New Guinea C strain) was amplified by PCR, comprising Domain III (amino acids 295 to 395) of the E protein. The fragment was cloned into pMD18-T vector and subcloned to expression vector pET-28a and pMAL-c2X. The recombinant plasmid pET-28a-D2EIII was transformed into E.coli BL21(DE3) and the pMAL-c2X-D2EIII was transformed into E.coli TB1. The induced recombinant proteins were purified by His-tag and MBP-tag affinity chromatography, respectively. The purified protein His-D2EIII was used to immunize rabbit three times at two-week intervals, the immunized rabbit produced high titer anti-His-D2EIII polyclonal antibody. The result of western blot indicated that the expressed fusion protein could react with the polyclonal antibody against Domain III of E protein.
The gene fragment coding for amino acids 281 to 395 of the E protein of DENV-2 （New Guinea C strain） was amplified by PCR, comprising Domain Ⅲ （amino acids 295 to 395） of the E protein. The fragment was cloned into pMD18-T vector and subcloned to expression vector pET-28a and pMAL-c2X. The recombinant plasmid pET-28a-D2EⅢ was transformed into E.coli BL21（DE3） and the pMAL-c2X-D2EⅢ was transformed into E.coli TB 1. The induced recombinant proteins were purified by His-tag and MBP-tag affinity chromatography, respectively. The purified protein His-D2EⅢ was used to immunize rabbit three times at two-week intervals, the immunized rabbit produced high titer anti-His-D2EⅢ polyclonal antibody. The result of western blot indicated that the expressed fusion protein could react with the polyclonal antibody against Domain III of E protein.
出处
《中国病毒学》
CSCD
2006年第6期619-621,共3页
Virologica Sinica
基金
国家"863"高技术发展计划资助项目(2005AA219070)
