嗜肺军团菌PAL抗原基因的克隆及真核表达  

Cloning and Expression of PAL Gene of Legionella pneumophila

作　　者：田玉[1] 陈建平[1] 杨春蕾[2] 刘明杰[1] 

机构地区：[1]四川大学华西基础医学与法医学院寄生虫学教研室,成都610041 [2]四川大学生命科学院医学细胞生物学教研室

出　　处：《四川大学学报（医学版）》2007年第4期583-586,598,共5页Journal of Sichuan University(Medical Sciences)

基　　金：教育部博士学科点专项科研基金(No.20060610091);四川省学术技术带头人培养基金(No.4200316)资助

摘　　要：目的构建真核表达重组质粒pcDNA3.1-PAL,并观察其在NIH3T3细胞中的表达。方法PCR扩增嗜肺军团菌PAL基因,定向克隆入真核表达载体pcDNA3.1(+)中,将经酶切、PCR扩增及序列测定正确的重组质粒命名为pcDNA3.1-PAL。脂质体法将重组质粒pcDNA3.1-PAL转染NIH3T3细胞,经免疫荧光和RT-PCR检测其瞬时表达和稳定表达产物。结果重组质粒pcDNA3.1-PAL转染NIH3T3细胞后获得了有效表达。结论成功构建了真核表达重组质粒pcDNA3.1-PAL为嗜肺军团菌核酸疫苗的进一步研究奠定了基础。Objective To develop DNA vaccine for Legionella pneumophila. Methods PAL gene of Legionella pneumophila was amplified with PCR. The amplified DNA was ligated to peDNA3.1 （＋） vector. The recombinant plasmid, which was identified by restriction analysis, PCR and sequence analysis, was named peDNA3. 1-PAL. The NIH3T3 cells were transfected with the recombinant plasmid peDNA3. 1-PAL by Lipofecti. Transient and stable expression products of the PAL gene were detected by immunofluorescence and RT-PCR. Results The recombinant plasmid pcDNA3. 1-PAL expressed PAL protein in the eukaryotic cell NIH3T3. Conclusion This study has built a foundation for the development of PAL gene DNA vaccine for Legionella pneumophila.

关 键 词：嗜肺军团菌 PAL 基因克隆 基因表达 

分 类 号：Q78[生物学—分子生物学]