中文摘要：

通过DNA重组技术表达沙眼衣原体（Chlamydia trachomatis，Ct）多形态膜蛋白D（polymorphic membrane protein D，PmpD），用Ct-PmpD重组蛋白（rCt—PmpD）制备免疫兔血清，观察该免疫血清在鸡胚攻毒试验中的保护作用。采用PCR技术从Ct L2型基因组中扩增PmpD基因，连接pET32a（＋）表达载体，转化宿主细胞E.coli BL21（DE3），IPTG诱导表达rCt-PmpD，用Ct L2免疫兔血清Western Blot（WB）检测其抗原性。复性后的rCt-PmpD免疫家兔制备免疫血清，免疫荧光法（IF）检测其免疫原性，并用鸡胚攻毒试验检测其中和活性。rCt-PmpD在工程菌中获得高效表达，以包涵体的形式存在胞浆中，WB结果显示rCt—PmpD能被ctL2免疫兔血清识别。用rCt-PmpD免疫家兔获得的血清在IF中能与Ct L2反应；中和试验表明，rCt—PmpD免疫血清能有效保护鸡胚免于死亡。成功构建重组表达载体rpET32a—PmpD，表达的rCt—PmpD具有良好的免疫原性，rCt—PmpD免疫血清具有中和活性，为进一步研制亚单位疫苗奠定基础。

英文摘要：

To clone and express the PmpD gene from Chlamydia trachomatis by DNA recombinant technology, analyze the immunogenicity of the recombinant protein, and identify neutralization of its antiserum. The PmpD gene was amplified by PCR with the genomic DNA from Chlamydia trachomatis serovar L2 as template. The amplified product was cloned into pET-32a（ ＋ ） vector, then transferred into the host cells E. coli BI21 （ DE3 ） strain. The recombinant PmpD was induced by IPTG, then purified, denatured and renatured. Its antigenicity was analyzed by Western-blotting with Ct L2 anti-rabbit serum. The serum was prepared after immuning the rabbits, and then its immunoreactivity was detected by immumofluorescence method. The recombinant PmpD was expressed with inclusion body in E. coli. Immunoblotting and Western-blotting revealed that recombinant protein had specific reaction with Ct L2 polyclonal antibody. The high antibody titer is detected in rabbit serum, and the serum can recognize Ct L2 specifically. The neutralization test indicate that the eggs are effectively protected by anti-rCt-PmpD rabbit serum. The recombinant expression plasmid pET-32a-PmpD is successfully constructed. The recombinant protein is of the good antigenicity, and the anti-rCt-PmpD polyclonal antibody can neutralize the Ct L2 infection. These results may provide the foundation for the further development of PmpD subunit vaccine.