嗜肺军团菌PAL、HSP60抗原基因在真核细胞中表达的实验研究  

Experimental Study of Expression of PAL Gene and HSP60 Gene of Legionella Pneumophila in NIH3T3 Cell

作　　者：田玉[1] 陈建平[1] 张建国[2] 杨春蕾[3] 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 [2]四川大学华西基础医学与法医学院形态学实验室,四川成都610041 [3]四川大学生命科学院医学细胞生物学教研室,四川成都610041

出　　处：《航天医学与医学工程》2007年第3期188-192,共5页Space Medicine & Medical Engineering

基　　金：高等学校博士学科点专项科研基金(No20060610092);四川省学术技术带头人培养基金(No4200316)资助

摘　　要：目的研究嗜肺军团菌PAL、HSP60抗原基因在真核细胞中的表达,为嗜肺军团菌DNA疫苗的研制提供实验依据。方法将本室已构建的重组质粒pcDNA3.1-PAL和重组质粒pcDNA3.1-HSP60分别及混合转染NIH3T3细胞,通过免疫荧光和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、pcDNA3.1-HSP60以及混合pcDNA3.1-PAL和pcDNA3.1-HSP60转染NIH3T3细胞后均获得了有效表达。结论本研究结果为嗜肺军团菌核酸疫苗的进一步研究奠定了基础。Objective To study the expression of PAL,HSP60 gene of Legionella pneumophila in NIH3T3 cell,in order to provide experimental data for developement of DNA vaccine against legionnaires＇ disease. Methods Recombinant plasmid pcDNA3.1-PAL and pcDNA3.1-HSP60 were transfected into the eukaryotic cell NIH3T3 respectively with lipofection strategy. Furthermore, mixed pcDNA3.1- PAL and pcDNA3.1-HSP60 were also transfected into NIH3T3. Their expression were detected by immunofluorescent staining and RT-PCR. Results Immunofluorescent staining and RT-PCR detection showed that the recombinant plasmid pcDNA3.1-PAL and pcDNA3.1-HSP60 could express PAL protein and HSP60 protein in the eukaryotic cell NIH3T3 respectively, and mixed pcDNA3.1-PAL and pcDNA3.1-HSP60 could express PAL protein and HSP60 protein respectively in the eukaryotic cell NIH3T3. Conclusion These evidences could be the basis for developing new genetic vaccine against Legionnaires＇ disease.

关 键 词：嗜肺军团菌 肽聚糖关联脂蛋白 热休克蛋白60 真核表达 

分 类 号：Q789[生物学—分子生物学]