全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

嗜肺军团菌mip基因重组质粒GFP-mip的构建及表达

Keywords: 嗜肺军团菌,mip基因,GFP-mip,蛋白表达

Full-Text   Cite this paper   Add to My Lib

Abstract:

目的构建嗜肺军团菌mip基因的真核重组质粒GFP-mip,并观察其在NIH3T3细胞中的表达。方法以嗜肺军团菌DNA为模版,通过PCR扩增获得mip基因,将其定向克隆到绿色荧光质粒pEGFP-C1中,构建真核重组质粒GFP-mip。经限制性核酸内切酶XhoI和BamHI酶切鉴定、PCR和核酸序列分析后,通过脂质体法转染到NIH3T3细胞中,利用荧光显微镜观察重组质粒的稳定表达。结果扩增出了702bpmip基因,在细胞质和细胞膜观察到较强绿色荧光。结论成功构建了真核重组质粒GFP-mip,并在NIH3T3细胞中得到了表达。

References

[1]  刘洪亮,陈学敏.嗜肺军团菌的研究进展(一)[J].中国卫生工程学,2006,5(2):109-112.
[2]  Barry S.Fields,Robert F.Benson,Richard E.Besser.Legionella and Legionnaires' disease: 25 years of investigation[J]. Clin Microbiol,2002,15(3):506-526.
[3]  Andreas Ceymann,Martin Horstmann,Philipp Ehses,et al.Solution structure of the Legionella pneumophila Mip-rapamycin complex[J].BMC Structural Biology,2008, 8(17):1-12.
[4]  JH Helbig,PC Luck,M Steinert, et al. Immunolocalization of the Mip protein of intracellularly and extracellularly grown Legionella pneumophila[J]. Microbiology,2003,32,83-88.
[5]  Wagner C,Khan AS,Kamphausen T,et al.Collagen binding protein Mip enables Legionella pneumophila to transmigrate through a barrier of NCI-H292 lung epithelial cells and extracellar matrix[J].Cellular Microbiology,2007, 9(2): 450-462.
[6]  Newton HJ, Ang DK, van Driel IR,et al. Molecular pat-hogenesis of infections caused by Legionella pneumophila[J]. Clin Microbiol Rev,2010,4(23):274-298.
[7]  Rolf Kohler,JorgFanghnel,BettinaKonig.Biochemical and Functional Analyses of the Mip Protein:Influence of the N-Terminal Half and of Peptidylprolyl Isomerase Activity on the Virulence of Legionella pneumophila[J].Infection and Immunity, 2003,8:4389-4397.
[8]  Alan Riboldi-Tunnicliffe, Bettina Konig, Sven Jessen, et al. Crystal structure of Mip, a prolylisomerase from Legionella pneumophila[J]. Nature structural boilogy,2001,9(8):779-783.
[9]  JurgenH.Helbig,Bettina Konig,Henry Knospe, et al.The PPIase Active Site of Legionella pneumophila Mip protein is involved in the infection of Eukaryotic host cells[J]. Biology Chemistry, 2003,384(1):450-462.
[10]  邵祝军, 万超群, 徐建国.军团菌巨噬细胞感染增强蛋白研究及其应用前景[J].中华流行病学杂志,2001,4(2),146-148.
[11]  Wang T, CHEN JP, LI H.Co-expression and Immunity of Legionella pneumophila mip Gene and Immunoadjuvant ctxB Gene[J].Biochemistry and Cell Biology,2005,37(3): 199-204.
[12]  Emre Dikici,Sylvia Daunert. Fluorescent timers shine a light on protein trafficking[J]. Nature Chemical Biology,2009, 2(5): 70-71.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133