| 赖型钩端螺旋体外膜蛋白LipL32基因真核表达载体的构建及表达的初步研究 |
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| 引用本文： | 黄毕,鲍朗,钟琪,商正玲,张会东,张英. 赖型钩端螺旋体外膜蛋白LipL32基因真核表达载体的构建及表达的初步研究[J]. 细胞与分子免疫学杂志, 2008, 24(2): 109-112 |
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| 作者姓名： | 黄毕 鲍朗 钟琪 商正玲 张会东 张英 |
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| 作者单位： | 四川大学华西基础医学与法医学院感染免疫研究室,四川,成都,610041 |
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| 基金项目： | 国家自然科学基金 , 教育部高等学校博士学科点专项科研基金 |
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| 摘 要： | 目的:构建赖型钩端螺旋体外膜蛋白LipL32基因真核表达载体并在COS-7细胞中表达,为钩端螺旋体DNA疫苗的研究和开发奠定基础.方法:从赖型钩端螺旋体017株全基因组中PCR扩增出目的基因,双酶切构建重组质粒pcDNA3.1-LipL32.脂质体转染法将重组质粒转染COS-7细胞,通过RT-PCR、Western blot检测目的基因的表达.结果:成功构建了LipL32基因的真核表达载体,并在COS-7细胞中获得瞬时和稳定表达.结论:赖型钩端螺旋体外膜蛋白LipL32基因真核表达载体能在哺乳动物细胞内表达,为钩端螺旋体DNA疫苗的应用提供了实验依据.
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| 关 键 词： | 钩端螺旋体 外膜蛋白LipL32 真核表达 赖型 钩端螺旋体 外膜蛋白 基因真核表达载体 构建及表达 研究 Leptospira gene outer membrane protein expression recombinant vector 实验 应用 胞内表达 哺乳动物 体能 稳定表达 结果 检测目 Western |
| 文章编号： | 1007-8738（2008）02-0109-04 |
| 收稿时间： | 2007-05-16 |
| 修稿时间： | 2007-06-18 |
Construction of the eukaryotic recombinant vector and expression of the outer membrane protein LipL32 gene from Leptospira serovar Lai |
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| HUANG Bi,BAO Lang,ZHONG Qi,SHANG Zheng-ling,ZHANG Hui-dong,ZHANG Ying. Construction of the eukaryotic recombinant vector and expression of the outer membrane protein LipL32 gene from Leptospira serovar Lai[J]. Chinese journal of cellular and molecular immunology, 2008, 24(2): 109-112 |
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| Authors: | HUANG Bi BAO Lang ZHONG Qi SHANG Zheng-ling ZHANG Hui-dong ZHANG Ying |
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| Affiliation: | Department of Infection and Immunity, Preclinical and Forensic Medicine School of West China Medical Center, Sichuan University, Chengdu 610041, China. |
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| Abstract: | AIM:To construct the eukaryotic experssion vector of LipL32 gene from Leptospira serovar Lai and express the recombinant plasmid in COS-7 cell. METHODS: The LipL32 gene was amplified from Leptospira strain 017 genomic DNA by PCR and cloned into pcDNA3.1, through restriction nuclease enzyme digestion. Then the recombinant plasmid was transformed into E.coli DH5alpha. After identified by nuclease digestion, PCR and sequencing analysis, the recombinant vector was transfected into COS-7 cell with lipsome. The expression of the target gene was detected by RT-PCR and Western blot. RESULTS: The eukaryotic experssion vector pcDNA3.1-LipL32 was successfully constructed and stably expressed in COS-7 cell. CONCLUSION: The eukaryotic recombinant vector of outer membrane protein LipL32 gene from Leptospira serovar Lai can be expressed in mammalian cell, which provides an experimental basis for the application of the Leptospira DNA vaccine. |
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| Keywords: | Leptospira outer membrane protein LipL32 eukaryotic expression |
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