期刊论文

Prokaryotic expression of the capsid protein VP1 of Chlamydiapsittaci bacteriophage Chp1 and preparation of its polyclonal antibodies

中文

中国病原生物学杂志

1673-5234

2018

13

9

975-978,983

10.13350/j.cjpb.180910

湖南省自然科学基金项目（No.2016JJ3103） 湖南省教育厅科学研究项目（No.16C1390）

本校为第一机构

目的原核表达并纯化鹦鹉热衣原体(Chlamydia psittaci,Cps)噬菌体Chp1衣壳蛋白VP1的重组蛋白rVP1_(1-190),并制备多克隆抗体。方法采用Clustal Omega在线软件对6株衣原体噬菌体VP1蛋白的氨基酸序列进行多重比对,确定Chp1VP1蛋白的保守区和特异区,并用DNAStar软件预测抗原表位。选择VP1_(1-190)与pET28a(+)载体连接构建原核表达载体pET28a-VP1_(1-190),转化至E.coli BL21(DE3)中诱导表达,采用Ni-NTA亲和层析法纯化重组蛋白;用纯化的重组蛋白免疫ICR小鼠,制备免疫血清,ELISA法检测其抗rVP1_(1-190)的效价。结果构建的原核表达载体pET28(a)-VP1_(1-190)经IPTG诱导高效表达了相对分子质量约为30×10...

Objectives To express the recombinant protein rVP1_(1-190) of Chlamydia psittaci phage Chp1 capsid protein VP1 in a prokaryotic expression system,to purify that protein,and to prepare polyclonal antibodies against it. Methods The online software Clustal Omega was used to align the amino acid sequences of capsid protein VP1 from 6 chlamydiaphages and to determine the conserved and specific regions of Chp1 capsid protein VP1.The software DNAStar was used to predict its antigenic epitopes.VP1_(1-190) was ligated into pET-28 a(+)to construct the pr...