肺炎衣原体假定蛋白Cpn0146的定位及其生物信息学分析

Localization and bioinformatics analysis of Chlamydial pneumonia hypothetical protein cpn0146

目的:对肺炎衣原体假定蛋白Cpn0146进行定位与生物信息学分析。方法:使用PCR方法从肺炎衣原体菌株AR39基因组中扩增Cpn0146,并将其插入到pGEX-6P2构建表达质粒pGEX-6p2-Cpn0146并转化大肠杆菌XL-Blue,IPTG诱导其表达融合蛋白GST-Cpn0146。纯化后的融合蛋白免疫小鼠制备特异性抗体,并应用间接免疫荧光法对肺炎衣原体的Cpn0146进行定位分析,最后利用软件Expasy和antheprot5.0对Cpn0146进行生物信息学分析。结果:成功构建了表达载体pGEX-6p2-Cpn0146和制备了抗Cpn0146的多克隆抗体。间接免疫荧光实验显示Cpn0146位于肺炎衣原体的包涵体膜上。生物信息学分析显示Cpn0146具有较好的亲水性与抗原性。结论:肺炎衣原体假定蛋白Cpn0146为一包涵体膜蛋白,其抗原性和亲水性较好。

Objective：To perform localization and bioinformatics analysis on Chlamydial pneumonia hypothetical protein cpn0146.Method：Cpn0146 was amplified from Chlamydia pneumonia AR39 genome and inserted to vector pGEX-6P2 to recombine expression plasmid pGEX-6P2-cpn0146.Then E.Coli XL-blue transformed with pGEX-6P2-cpn 0146 was induced with IPTG to express fusion protein GST-cpn0146.The purified GST-CPN0146 was used to immunize mice and indirect immunofluorescence assay（IFA）was adopted to carry out localization analysis of cpn0146.Finally bioinformatics analysis of Chlamydial pneumonia hypothetical protein cpn0146s was performed with software Expasy and antheprot 5.0..Result：pGEX-6P2-cpn0146 was constructed successfully and anti-Cpn0146 polyclonal antibody was made successfully.The hypothetical protein Cpn0146 was demonstrated to locate in the inclusion membrane of Chlamydia pneumonia using IFA.Furthermore,bioinformatics analysis showed Cpn0146 had good hydrophobicity and antigenicity.Conclusion：Chlamydial pneumonia hypothetical protein cpn0146 was an inclusion membrane protein and had good hydrophobicity and antigenicity.