摘要
对含伪狂犬病病毒 ( PRV)鄂 A株 g G全基因 ,PK、g D基因部分编码序列的质粒 p USK进行改造 ,消除其中的 Eco R 位点 ,将通过 PCR扩增的增强绿色荧光蛋白基因 ( EGFP)融合到 g G启动子下游 ,构建了由 g G启动子控制 EGFP表达的 PRV转移载体 pg G- /EGFP+。该转移载体单独转染或同 PRV基因组 DNA共转染 IBRS-2细胞 ,结果单独转染时 EGFP不能表达 ;共转染时 ,6 h就可在荧光显微镜下观察到明显的荧光。
The EcoRⅠ site of plasmid pUSK containing the complete gG gene, partial PK and gD genes, was removed to result in the plasmid pUSK△E. The coding sequence of enhanced green fluorescent protein (EGFP) was amplified from pEGFP-C1 by PCR and cloned into pUSK△E under the control of the gG promoter, to obtain the transfer vector pgG -/EGFP +. When pgG -/EGFP + was transfected into IBRS-2 cells alone or cotransfected with the genome DNA of pseudorabies virus Ea strain, it is interesting that the cells which transfected pgG -/EGFP + alone could not emit green fluoresence; whereas, a green fluoresence could be observed in the cotranfected cells and as early as 6 h post-transfection.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2001年第2期125-127,共3页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目! (39970 5 5 9)
