摘要
为了构建微小隐孢子虫子孢子表面蛋白CP2 3基因真核表达载体pCR3 1 2 3,观察其在Hela细胞中的表达。本研究用EcoRⅠ从pMD18 T 2 3中酶切得到CP2 3基因片段 ,将其插入真核表达载体pCR3 1(+)的EcoRⅠ位点 ,构建CP2 3基因真核表达载体pCR3 1 2 3,脂质体介导法将其转染Hela细胞 ,并用G4 18加压筛选 ,用RT PCR方法检测外源CP2 3基因的转录、ELISA法和间接免疫荧光法检测其活性。酶切鉴定表明已成功构建了重组真核表达载体pCR3 1 2 3;外源CP2 3基因能在转染细胞中有效转录 ;
To construct an eukaryotic expressing vector pCR3 1 23 and express it in Hela cells,CP23 gene of C.parvum was obtained from pMD18 T 23 digested with EcoR Ⅰ, and was inserted into eukaryotic expressing vector pCR3 1(+) at EcoR Ⅰsite.Then Hela cell lines were transfected with this recombinant plasmid by liposomes.The transcription and expressed products of CP23 in the tranfected Hela cells were assayed by RT PCR,ELISA and indiret immunofluorescence assay after screening with G418.The results showed that pCR3 1 23 was constructed successfully. CP23 gene was transcripted in transfectants,and CP23 protein with obvious biological activity was highly expressed in Hela cells.
出处
《寄生虫与医学昆虫学报》
CAS
2003年第1期16-20,共5页
Acta Parasitologica et Medica Entomologica Sinica
基金
吉林省杰出青年基金 (2 0 0 0 )
