阳离子脂质体介导日本血吸虫组织蛋白酶L1基因的真核表达  

Eukaryotic expression cathepsin L proteases of Schistosoma japonicum using cationic lipids as gene delivery vector

作　　者：刘金辉[1] 严涛[1] 黎帆[1] 余克花[1] 詹希美[2] 易冰[2] 

机构地区：[1]南昌大学医学院病原生物学教研室,南昌330006 [2]中山大学中山医学院寄生虫学教研室,广州510080

出　　处：《中国人兽共患病学报》2008年第1期63-66,共4页Chinese Journal of Zoonoses

摘　　要：目的真核表达日本血吸虫组织蛋白酶L1(SjCL1)以研究其生物学功能。方法采用阳离子脂质体复合质粒SjCL1/AD1-1 DNA并转染HeLa细胞,通过RT-PCR检测SjCL1基因在HeLa细胞中的转录,SDS-PAGE电泳法分析目的基因的蛋白表达产物,并进一步通过Western Blot法证实。结果RT-PCR检测到转染的细胞中有与目的基因相符大小约1000bp转录条带,SDS-PAGE电泳发现表达质粒SjCL1/AD1-1DNA转染的细胞上清液中存在大小约为31kDa的表达蛋白带,Western Blot法证实该表达蛋白产物可和日本血吸虫兔血清发生特异的反应。结论阳离子脂质体将SjCL1/AD1-1转染入HeLa细胞后可特异地表达一个大小约为31kDa的可分泌的日本血吸虫蛋白产物。In order to analyze the expression of cathepsin L proteases of Schistosoma japonicum （SiCL1） for the further study of its bioactivity, eukaryotic expression plasmid SjCL1/ADI-1 was constructed and transfected into HeLa cells, using cationic lipid as gene delivery vector. A transcript with size of about 1000bp was detected by RT-PCR in the tranfected cells. An expressed protein of 31kDa was found through SDS-PAGE, and the expressed protein could specifically react with rabbit serum against Schistosomajaponicum in as demonstrated in Werstern blotling. It suggests that SjCL1 gene could be expressed efficiently from HeLa cells for the analysis of its bioactivities, when cationic eipid was used as vector for gene delivery.

关 键 词：日本血吸虫 组织蛋白酶L 阳离子脂质体 基因转染 基因表达 

分 类 号：R383.2[医药卫生—医学寄生虫学]