沙眼衣原体隐匿质粒编码蛋白的表达及其相互作用  

Cryptic plasmid-encoded proteins in Chlamydia trachomatis: expression and interaction

作　　者：高磊琼 张燕[2] 华子瑜[1,2] 沈犁[2] 

机构地区：[1]重庆医科大学附属儿童医院新生儿科,重庆400014 [2]重庆医科大学附属儿童医院儿童发育疾病研究教育部重点实验室,儿科学重庆市重点实验室,重庆市儿童发育重大疾病诊治与预防国际科技合作基地,重庆400014

出　　处：《第三军医大学学报》2015年第1期34-38,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81370777);国家临床重点专科-新生儿学项目(卫办医政函[2011]873号)~~

摘　　要：目的分析沙眼衣原体隐匿质粒编码蛋白的相互作用。方法采用基因克隆的方法将沙眼衣原体质粒基因pgp1~4及pgp8分别克隆到pRAC和pRBR上,通过转录激活细菌双杂交系统分析蛋白相互作用。结果 Western blot结果显示Pgp1、Pgp3与大肠杆菌Rpo A氨基末端功能区（α-NTD）融合蛋白及Pgp1、Pgp3、Pgp4与DNA结合蛋白λCI融合蛋白,能在大肠杆菌中表达。在报告菌株中,共表达与α-NTD融合的Pgp3蛋白（α-Pgp3）及与λCI融合的Pgp3蛋白（CI-Pgp3）,导致报告基因产物β-半乳糖苷酶的活性升高。而余无明显变化。结论 pgp3编码的Pgp3能与自身相互作用。Objective To analyze the interactions among the proteins encoded in the cryptic plasmid of Chlamydia trachomatis （ C. trachomatis）. Methods The DNA fragments of C. trachomatis plasmid genes, pgpl ～ 4 and pgp8, were cloned into the vectors （pRAC and pRBR） , and the target genes were in-frame fused to α subunit N-terminal domain （ot-NTD） and DNA binding protein, CI protein of bacteriophage λ （λCI） , respectively. The expression of cloned genes in Escherichia coli（ E. coli） were determined by Western blotting. The protein-protein interactions were analyzed by a transcription-based bacterial two-hybrid system. Results Western blotting indicated that isopropyl β-D-1-thiogalactopyranoside （IPTG） induced the expression of fusion proteins, α-Pgpl, α-Pgp3, CI-Pgpl, CI-Pgp3 and CI-Pgp4 in E. coli. In the presence of Pgp3 （encoded by plasmid pBRot-Pgp3 ）, the expression of α-Pgp3 and CI-Pgp3 activated the transcription from a reporter gene β-galactosidase in the bacterial two-hybrid assay strain, but Pgp3 had no interaction with other test proteins. Conclusion Pgp3 encoded by pgp3 interacts with Pgp3 in C. trachomatis.

关 键 词：沙眼衣原体 隐匿质粒 Pgp3 蛋白相互作用 

分 类 号：R374.1[医药卫生—病原生物学] R394-33[医药卫生—基础医学]