鹦鹉热衣原体HSP60基因克隆及其表达产物抗原性的实验研究

Cloning and expression of gene encoding the heat shock protein-60 from Chlamydia psittaci and the immunogenicity analysis of its gene product

目的验证新获得的鹦鹉热衣原体HSP60基因在E.coli中高效表达的产物是否具有足够的抗原活性,为进一步研究其在鹦鹉热衣原体感染检测上的应用奠定基础。方法根据GenBank鹦鹉热衣原体Hsp60基因序列X51404,设计一对特异性引物,扩增了鹦鹉热衣原体HSP60全长基因,克隆入pET32a(+)载体,转化大肠杆菌BL21(DE3)后,利用IPTG诱导获得高效表达。结果扩增了鹦鹉热衣原体HSP60全长基因,表达产物分子量约为68kD,经Western blotting和胶体金免疫层析技术检测,表明表达产物具有较好的抗原性。结论扩增了鹦鹉热衣原体HSP60全长基因,并对其表达产物的抗原性进行免疫印记分析和免疫层析初步检测,表明具有较好的抗原性,为进一步研究其在鹦鹉热衣原体感染检测上的意义打下了基础。

To clone and express the gene encoding the heat shock protein-60 （Hsp-60） from Chlarnydia psittaci, and to analyze the immunogenicity of its gene product, the full length of the Hsp-60 gene was amplified with a specific primer designed from the Hsp-60 gene sequence X51404 of C. psittaci of GenBank and cloned into plasmid pET32a（ ＋ ）. High efficient expression of the gene was obtained after transforming into E. coli BL21 （DE3） and induction with IPTG. Purification of the gene product was carried out for the purpose to apply the recombinant protein. The experimental results showed that the molecular weight of the expressed product was 68 kDa and the result in Western blot analysis and colloidal gold immunochromatography indicated that the expressed protein showed a better and specific immunogenicity. It is concluded that the stable expression and the better and specific immunogenicity of the expressed gene product of C. psittaci provide foundation for the further studies on infection with C. psittaci.