摘要
利用构建的犬2型腺病毒E3区缺失质粒pBE3L分别构建了含有和不含外源性启动子的绿色荧光蛋白(GFP)基因和狂犬病病毒糖蛋白(Rgp)基因的重组表达质粒pBE3LGFP、pBE3LCGFP、pBE3LRgp和pBE3LCRgp,并分别对DK细胞进行了转染实验,以检测其表达,结果显示,pBE3LCGFP质粒在转染DK细胞后,于36h即可观察到荧光,72～96h无明显差别,传3代后仍可见表达荧光的细胞,pBE3LCRgp质粒转染DK细胞后,于48～96h用间接免疫荧光染色可检测到糖蛋白的表达,而pBE3LGFP和pBE3Rgp质粒转染DK细胞后经检测均无表达,表明构建的E3区缺失性载体不能利用E3区自身的启动子进行目的基因的表达,但利用外源性启动子可使外源基因获得良好表达。
Several expressing vectors with or without forei gn promoter were constructed on the basis of deleted E3region of CAV-2using the GFP gene and Rgp gene as the report genes.The vectors were transfected into th e DK cells to de-tect their expression.The results showed that the vectors wit h foreign promoter can express the report genes,while the vectors without fore ign promoter can not express the report genes.
出处
《生物技术通讯》
CAS
2003年第1期8-11,15,共5页
Letters in Biotechnology
基金
全军"十五"医药卫生重点项目(01-Z-092)
国家自然科学基金项目(30170704)
