摘要
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。
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 eukaryotic expression 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 into Vero cell by LipofectamineTM2000. After 48 hours, green fluorescent could be seen in Vero cells under fluorescence microscope. This fusion gene was highly expressed in Veto cells. It has laid a good foundation for studying further the biological ftmction of heat shock protein.
出处
《吉林畜牧兽医》
2012年第4期31-33,共3页
Jilin Animal Husbandry and Veterinary Medicine
基金
国家自然科学基金项目(30972176)
关键词
猪囊尾蚴
热休克蛋白35.6
真核
质粒
Cysticercus cellulosae
heat shock protein 35.6
eukaryotic
plasmid
