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沙眼衣原体CT-249基因编码蛋白为一包涵体膜蛋白
引用本文:贾天军,刘殿武,罗建华,钟光明.沙眼衣原体CT-249基因编码蛋白为一包涵体膜蛋白[J].微生物学报,2007,47(4):645-648.
作者姓名:贾天军  刘殿武  罗建华  钟光明
作者单位:1. 河北医科大学流行病学教研室,石家庄,050017
2. Department of Microbiology and Immunology, University of Texas Health Science Center at San Antonio,7703 Floyd Curl Drive,San Antonio, Texas 78229,USA
摘    要:使用融合蛋白GST-CT249的抗体对假想蛋白CT249的特性进行研究。使用PCR方法从L2型沙眼衣原体的基因组中扩增编码CT249蛋白的开放读码区基因,限制性内切酶BamHⅠ和NotⅠ消化、T4连接酶连接导入pGEX-6p2载体,进一步把重组质粒pGEX-6p2-CT249转化到XL1-blue细菌,并诱导表达融合蛋白GST-CT249。在融合蛋白GST-CT249免疫小鼠制备抗体后,应用直接免疫荧光技术对衣原体感染细胞内的CT249基因表达的内源性蛋白进行初步定位。成功克隆出沙眼衣原体基因CT249,全长为351bp,并表达了融合蛋白GST-CT249,分子量为38.2kDa。制备了融合蛋白GST-CT249的抗体并初步定位假想蛋白CT249于沙眼衣原体包涵体膜蛋白上。总之,使用融合蛋白GST-CT249的抗体,鉴定假想蛋白CT249为一种新的沙眼衣原体包涵体膜蛋白。该发现将为进一步深入研究衣原体与宿主细胞间某些机制提供了有用的途径。

关 键 词:沙眼衣原体  融合蛋白  克隆  表达  包涵体膜蛋白
文章编号:0001-6209(2007)04-0645-04
收稿时间:2006/11/1 0:00:00
修稿时间:2/5/2007 12:00:00 AM

Localization of the hypothetical protein CT249 in the Chlamydia trachomatis inclusion membrane
JIA Tian-jun,LIU Dian-wu,LUO Jian-hua and ZHONG Guang-ming.Localization of the hypothetical protein CT249 in the Chlamydia trachomatis inclusion membrane[J].Acta Microbiologica Sinica,2007,47(4):645-648.
Authors:JIA Tian-jun  LIU Dian-wu  LUO Jian-hua and ZHONG Guang-ming
Institution:Department of Epidemiology; Hebei Medical University; Shijiazhuang 050017; China;Department of Epidemiology; Hebei Medical University; Shijiazhuang 050017; China;Department of Microbiology and Immunology; University of Texas Health Science Center at San Antonio; 7703 Floyd Curl Drive; San Antonio; Texas 78229; USA;Department of Microbiology and Immunology; University of Texas Health Science Center at San Antonio; 7703 Floyd Curl Drive; San Antonio; Texas 78229; USA
Abstract:To characterize the hypothetical protein CT249 using antibodies raised with CT249 fusion protein. The open reading frame (ORF) coding for CT249 in the Chlamydia trachomatis serovar L2 genome was cloned into the pGEX-6p2 vector using the restriction enzymes BamH I and Not I. The recombinant plasmid pGEX-6p2-CT249 was transformed into XL1-blue bacteria and the gene CT249 was expressed as fusion proteins with the glutathione-s-transferase (GST) tagged to the N-terminus. The GST-CT249 fusion protein was used to immunize mice and the mouse anti-fusion protein antibody was used to localize the endogenous CT249 protein in Chlamydia-infected cells using an indirect immunofluorescence assay (IFA). The CT249 gene with 351bps in length was successfully cloned and expressed as GST fusion protein with molecular weight of 38.2kDa. The anti-fusion protein antibodies produced from mice detected the hypothetical protein CT249 in the inclusion membrane of Chlamydia trachomatis-infected cells. Using antibodies raised with GST-CT249 fusion protein, the hypothetical protein CT249 have been identified as a Chlamydia trachomatis inclusion membrane protein. Given the potentially important role of inclusion membrane proteins in chlamydial interactions with host cells, this finding has provided a useful tool for further understanding the mechanisms of chlamydial intracellular parasitism.
Keywords:Chlamydia trachomatis  fusion protein  expression  inclusion membrane protein
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