重组A型产气荚膜梭菌α毒素保护性抗原的制备及初步免疫保护作用研究

Preparation of Alpha-toxin’s Protective Antigen of Clostridium perfringens Type A and Research for Its Primary Immunological Protective Function

诱导表达重组工程菌pBV cpa4 0 8后 ,将表达菌体超声破碎 ,上清经 80 %饱和硫酸铵一次沉淀 ,经透析 ,上凝胶过滤层析柱进行分离纯化 ,薄层凝胶扫描结果显示 ,纯化的蛋白纯度达 95 %以上 ;用纯化蛋白免疫昆明小鼠 ,以 1 0MLD1 0 0 腹腔进行攻击 ,被免疫小鼠获得了 10 0 %的保护。

Induced by 42℃,the recombinant engineering bacterial pBV/cpa408 was highly expressed. After having been pelleted by 80%(NH 4) 2SO 4 and dialysised, the expressed protein was isolated and purified by the gel filtration choromatography. Then according to an amount of 1.0mg/kg, the Kunming Mice(body weighted 18g) were immuned with the purified protein by intraperitoneal inoculation. One week after the first enhanced immunization, the Kunming Mice were attacked with an amount of 1.0MLD alpha_toxin, in which the eight mice immuned all survive and the control group all died. During the period of immunization, the titre of the mouse′s serum antibody was measured by ELISA. One week after the first immunization, the titre of the mice′s serum antibody was 1∶800,but that of one week after the first enhanced immunization reached to 1∶6400.