|本期目录/Table of Contents|

[1]刘胜泉,曹秀琴,杨志伟.嗜肺军团菌pip基因原核质粒的构建及其表达[J].宁夏医科大学学报,2012,(01):12-13,21.
 LIU Sheng-quan,CAO Xiu-qin,YANG Zhi-wei.Construction and Expression of Pip Gene of Legionella Pneumophila in Prokaryotic Cell[J].Ningxia Medical University,2012,(01):12-13,21.
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2012年01期
页码:
12-13,21
栏目:
论著
出版日期:
2012-01-30

文章信息/Info

Title:
Construction and Expression of Pip Gene of Legionella Pneumophila in Prokaryotic Cell
作者:
刘胜泉1 曹秀琴2 杨志伟1
1.宁夏医科大学基础医学院病原生物学与免疫学系,银川 750004; 2.宁夏医科大学生育力保持省部共建教育部重点实验室,银川 750004
Author(s):
LIU Sheng-quan1 CAO Xiu-qin2 YANG Zhi-wei1
1.Dept. of Pathogenic Biology and Immunology, Ningxia Med. Univ., Yinchuan 750004; 2.Key Lab. of Education in Fertility Preservation and Maintenase,Yinchuan 750004
关键词:
嗜肺军团菌 pip基因 表达 纯化
Keywords:
Legionella pneumophila pip gene expression purification
分类号:
R378
DOI:
-
文献标志码:
A
摘要:
目的 扩增嗜肺军团菌基因pip,构建重组质粒pET-pip并在原核系统中表达。 方法 采用PCR,从嗜肺军团菌基因组DNA中扩增出pip基因,将其定向克隆至原核表达载体pET-32a(+),构建重组质粒pET-pip,经限制性内切酶、PCR鉴定及测序分析后,转化BL21,IPTG诱导表达,并用SDS-PAGE进行鉴定。使用His-tag纯化重组蛋白PIP。结果 扩增出了嗜肺军团菌726bp的pip基因,构建的原核表达重组质粒pET-pip表达并纯化出了约46kD Trx-PIP的融合蛋白。结论 成功构建了嗜肺军团菌pip基因的原核重组质粒,并在大肠杆菌中得到了高效表达。
Abstract:
Objective To construct the recombinant plasmid pET-pip and to detect its expression in the prokaryotic cells BL21(DE3). Methods The pip gene was amplified by PCR from DNA of Legionella pneumophila, then inserted it into the prokaryotic expression vector pET-32a(+).The recombinant plasmid pET-pip was constructed. The recombinant plasmid was identified by restriction-endonuclease digestion, PCR and DNA sequencing analysis and transferred into BL21.The expression of pET-pip was induced with IPTG and the fusion protein was analyzed with SDS-PAGE. It was purified by His-tag. Results The pip gene of 726bp in length was amplified and the recombinant plasmid pET-pip was constructed.The PIP protein of approximately 46KD in size was expressed in E. coli and purified.Conclusion The pip gene of 726bp in length was successfully cloned and the recombinant plasmid pET-pip was constructed successfully and expressed efficiently.

参考文献/References:

[1] Parthuisot N, Binet M, Touron-Bodilis A, et al. Total and Viable Legionella pneumophila Cells in Hot and Natural Waters as Measured by Immunofluorescence-Based Assays and Solid-Phase Cytometry[J]. Appl Environ Microbiol,2011,77(17):6225-6232.
[2] Declerck P, Behets J, van Hoef V, et al. Detection of Legionella spp. and some of their amoeba hosts in floating biofilms from anthropogenic and natural aquatic environments[J]. Water Research,2007,41(14):3159-3167.
[3] Benin AL, Benson RF, Besser RE. Trends in legionnaires disease, 1980-1998: declining mortality and new patterns of diagnosis[J]. Clin Infect Dis, 2002, 35(9):1039-1046.
[4] Stout JE, Muder RR, Mietzner S, et al. Role of environmental surveillance in determining the risk of hospital-acquired legionellosis: a national surveillance study with clinical correlations [J]. Infect Control Hosp Epidemiol,2007,28(7):818-824.
[5] Helbig JH, Uldum SA, Luck PC, et al. Detection of Legionella pneumophila antigen in urine samples by the BinaxNOW immunochromatographic assay and comparison with both Binax legionella Urinary Enzyme Immunoassay(EIA)and Biotest Legionella Urin Antigen EIA[J]. Med Microbiol,2001,50(6):509-516.
[6] Chien M, Morozova I, Shi S, et al.The genomic sequence ofthe accidental pathogen Legionella pneumophila[J]. Science,2004,305(5692):1966-1968.
[7] Sosa NL,Alfonso LR,Gonzalez MJ,et al. Legionella pneumophila:extraction of the main protein from the outer membrane[J]. Rev Cubana Med Trop,2002,54(2):91-95.
[8] Yang ZW, Chen JP, Wang T,et al. Immune response and protective efficacy in BALB/c mice induced by DNA vaccine encoding immunogenic protein of Legionella pneumophila[J]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2007,23(3):209-212.

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备注/Memo

备注/Memo:
收稿日期:2011-06-08 基金项目:国家自然科学基金资助项目(30860264); 宁夏自然科学基金(NZ0992) 作者简介:刘胜泉(1984-),男,河南人,在读硕士研究生,从事病原生物学专业。 通信作者:杨志伟,教授,硕士研究生导师,从事感染免疫和自身免疫研究工作。
更新日期/Last Update: 2012-01-20