期刊论文

Localization of 10Putative Effector Proteins Secreted via TypeⅢ Secretion System of Chlamydia psittaci

中文

畜牧兽医学报

0366-6964

2020

51

6

1354-1361

10.11843/j.issn.0366-6964.2020.06.018

国家自然科学基金(31600150) 湖南省大学生研究性学习与创新性实验计划项目(张旭) 湖南省教育厅一般项目(18C0457)。

本校为第一机构

旨在分析预测的10种鹦鹉热衣原体(Chlamydia psittaci,Cps)Ⅲ型分泌系统效应蛋白在细胞内的定位,为下一步研究其结构与功能奠定基础。本研究利用计算机辅助的生物信息学方法(Effective T3和BPBAac tool)预测Cps T3SS效应蛋白。利用分子克隆技术构建重组质粒pGEX-6P-1/目的基因,诱导表达、纯化相应重组蛋白。皮下免疫BALB/c小鼠,制备各重组蛋白的多克隆抗体,ELISA测定其效价。以制备的多克隆抗体为一抗,应用间接免疫荧光法检测假定的效应蛋白在感染细胞中的定位。本研究通过应用Effective T3、BPBAac tool交叉验证,预测了14个Cps T3SS效应蛋白编码基因:CPSIT_0357、CPSIT_0429、CPSIT_0461、CPSI...

This study aimed to investigate the intracellular localization of 10 putative effector proteins via typeⅢsecretion system(T 3 SS)in Chlamydia psittaci,laying the basis for the next structure and function analysis.In this study,computer-aided bioinformatics methods such as Effective T3 and BPBAac were used to select the possible T 3 SS effector proteins of C psittaci.The recombinant plasmid pGEX-6P-1-target genes were constructed and transformed into E.coli BL21,and the recombinant proteins were induced and purified.Then BALB/c mice were immuni...