恶性疟原虫HT1基因荧光真核表达载体的构建及细胞内表达  

Construction and cellur expression of the gene-fluorescence in eukaryotic expression vector of Plasmodium falciparum

作　　者：徐劲[1] 余新炳[1] 叶苓[1] 陆家海[1] 

机构地区：[1]中山医科大学寄生虫学教研室,广州510089

出　　处：《中国人兽共患病杂志》2002年第2期66-68,共3页Chinese Journal of Zoonoses

基　　金：国家"九五"攻关项目 (96 90 6 0 4 0 6);广东省卫生厅青年基金 (98 3 3 2 )资助

摘　　要：目的　体外扩增恶性疟原虫海南分离株的已糖转运体基因 (pfHT1) ,构建恶性疟原虫HT1基因真核融合表达载体并检测其表达情况 ,为用于DNA免疫作准备。方法　特定PCR引物的设计 ,扩增 ,基因组DNA提取 ,连接 ,鉴定 ,细胞培养 ,脂质体转染 ,荧光显微镜观察。结果　从恶性疟原虫海南分离株基因组DNA中扩增特异的编码 pfHT1的基因序列 ,片断大小为 15 16bp ,经酶切鉴定证实成功构建 pEGFP -HT1融合表达质粒 ,将其转染体外培养的细胞后 ,可见融合蛋白表达产生的明亮的绿色荧光。结论　成功构建 pEGFP -HT1融合表达质粒 ,并实现在细胞内的表达。为进行动物体内的DNA免疫打下了基础。Aim Amplifing and cloning the gene coding pfHT1 of Plasmodium falciparum for constructing eukaryotic fuse expression vector to study DNA viccine.Method The primers were designed following the sequence of pfHT1.In vitro culture of matarias,genome DNA distilled,PCR,DNA linking,transfection technique,construction of eukaryotic express plasmid etc were used in this thesis.Results The fragment of pfHT1 gene with 1516bp was amplified and then the recombination eukaryotic expression plasmid pEGFP HT1 was constructed by directional cloning.fluoresecnce of fuse protein expression was found in cells in vitro.Conclusion The reconbination eukarytic expression plasmid of pfHT1 gene was constructed successfully,and it has given the basic material for studying the structure and functions of pfHT1 genes.

关 键 词：绿色荧光蛋白 DNA疫苗 基因转染 恶性疟原虫 已糖转运体 基因表达 真核表达载体 

分 类 号：R382.31[医药卫生—医学寄生虫学]