| 嗜肺军团菌PAL抗原基因的克隆及真核表达 |
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| 引用本文： | 田玉,陈建平,杨春蕾,刘明杰.嗜肺军团菌PAL抗原基因的克隆及真核表达[J].四川大学学报(医学版),2007,38(4):583-586,598. |
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| 作者姓名： | 田玉 陈建平 杨春蕾 刘明杰 |
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| 作者单位： | 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;四川大学生命科学院,医学细胞生物学教研室 |
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| 基金项目： | 高等学校博士学科点专项科研项目
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四川省学术与技术带头人培养基金 |
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| 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.结论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.
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| 关 键 词： | 嗜肺军团菌 PAL 基因克隆 基因表达 |
| 修稿时间： | 2007-01-022007-04-17 |
Cloning and Expression of PAL Gene of Legionella pneumophila |
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| TIAN Yu,CHEN Jian-ping,YANG Chun-lei,LIU Ming-jie.Cloning and Expression of PAL Gene of Legionella pneumophila[J].Journal of West China University of Medical Sciences,2007,38(4):583-586,598. |
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| Authors: | TIAN Yu CHEN Jian-ping YANG Chun-lei LIU Ming-jie |
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| Institution: | Department of Parasitology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China |
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| Abstract: | Objective To develop DNA vaccine for Legionella pneumophila.Methods PAL gene of Legionella pneumophila was amplified with PCR.The amplified DNA was ligated to pcDNA3.1( )vector.The recombinant plasmid,which was identified by restriction analysis,PCR and sequence analysis,was named pcDNA3.1-PAL.The NIH3T3 cells were transfected with the recombinant plasmid pcDNA3.1-PAL by Lipofection.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. |
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| Keywords: | Legionella pneumophila Peptidoglycan-associated lipoprotein(PAL)Clone |
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