重组猪囊尾蚴热休克蛋白35．6真核质粒的构建及鉴定

为了构建猪囊尾蚴（Cysticercuscellulosae）热休克蛋白的真核表达载体，试验利用PCR技术扩增HSP35．6的基因，将其与增强型绿色荧光蛋白的基因先后连接到pVAXI真核表达载体上，得到pVAXI—HSP35．6-EGFP的重组质粒，经酶切及测序鉴定正确。采用脂质体介导的DNA转染法，将阳性重组质粒转染Vero细胞。转染48h后荧光显微镜下观察到明亮的绿色荧光，说明融合基因可在Vero细胞中高效表达。本研究为进一步研究该蛋白生物学功能奠定了基础。

Construction and Identification of Recombinant Eukaryotic Plasmid of Cysticercus Cellulosae Heat Shock Protein 35.6

In order to construct a eukaryotic expression vector of Cysticercus cellulosae heat shock protein 35.6.We combined HSP35.6 gene to enhanced green fluorescent protein gene with PCR technology,and then connected it to eukaryoticexpression vector pVAXI.It showed that the length and sequence of recombinant pVAXI-HSP35.6-EGFP plasmid by restriction enzyme digestion and sequencing were correct The selected positive recombinant plasmid was transfected intoVero cell by LipofectamineTM2000.After 48 hours,green fluorescent could be seen in Vero cells under fluorescencemicroscope.This fusion gene was highly expressed in Vero cells.It has laid a good foundation for studying further the biological function of heat shock protein.