摘要
设计1对引物对鸡败血支原体R株mga0553基因进行PCR扩增,并通过双酶切定向克隆到原核表达载体pET-32a(+),获得重组质粒pET-32a-mga0553。重组质粒经诱导表达并取表达产物进行分析,结果显示,mga0553基因获得了融合表达。Western-blotting证实,表达产物MGA_0553与MG阳性血清具有免疫反应性;生长抑制试验表明,鼠抗MGA_0553血清对MG细胞的增殖具有抑制作用;间接免疫荧光染色检测结果显示:MG细胞呈现较强的绿色荧光。研究结果表明mga0553基因编码蛋白MGA_0553是MG的一种膜相关蛋白。
A pair of primers was designed in order to amplify the mga0553 gene from R strain of Mycoplasma gallisepticum. To obtain recombinant plasmid pET-32a,the gene fragment were inserted into pET-32a vector after cleavage by corresponding enzymes. The recombinant plasmid were transformated into E.coli BL21 （DE3） competence ceils and induced with IPTG. The expressed products MGA_0553 were recognized by MG positive serum in Western-blotting test. The analysis results of anti-MGA_0553 serum on MG growth suggested that the serum could inhibit the growth of MG. In indirect fluorescence assay （IFA）,the result showed that mga0553 coding protein was positive on the MG cells. In conclusion, the mga0553 gene might encode a kind of membrane-assoicated proteins in MG.
出处
《中国家禽》
北大核心
2008年第24期18-21,共4页
China Poultry
基金
国家自然科学基金(30871883)
