羊种布氏杆菌核酸疫苗的制备及其免疫效果

Preparation nucleic acid vaccine of brucella melitensis and its immune effect

目的：构建羊种布氏杆菌基因组DNA表达文库，并研究其免疫效果。方法：将羊布氏杆菌基因组DNA经限制性内切酶 Hind Ⅲ消化后，克隆人 pSV－β-Gal质粒，构建合羊布氏杆菌基因组 DNA酶切片段的表达文库，用构建的表达文库经股四头肌免疫小鼠。结果：从文库中筛选了26个含羊种布氏杆菌基因组DNA片段的细菌克隆；免疫小鼠，可以刺激小鼠产生羊种布氏菌凝集抗体，并使淋巴细胞转化率明显增高。结论：构建的羊布氏杆菌基因组DNA表达文库可以刺激小鼠产生体液免疫应答和细胞免疫应答，此工作为制备有效的布氏杆菌及其它胞内寄生菌疫苗提供了实验基础。

Abstract Objective:To construct bmcella melitensis Br.melitensis genomic DNA expression library and study its immune effect.Meth-ods: Hind Ⅲ digested Br.melitensis genomic DNA fragments were cloned into pSV-β-Gal plasmds.The recombinant plasmids were transformedinto JM109 host bacteria. After large-scale isolation, the recombinant plasmids were inoculated into the quadriceps muscle of mice. In the 9thday after third boost immunization .agglutination antibodies against Br. melitensis and lymphocyte mitogenic ability of mice were determined. Re-sults : Expression library nucleic acid vaccine can induce the production of agglutination antibodies against Br. melitensis and stimulate the lym-phocyte porliferation primed by ConA. Conclusion:The genomic DNA expression library may provide a new vaccine against Br. melitensis infec-tion.