摘要
应用PCR技术直接从破伤风芽孢杆菌质粒DNA中扩增出 1 4Kd的破伤风毒素C片段基因 ,经DNA序列测定证明其为破伤风片段C基因。将此基因克隆入大肠杆菌谷胱甘肽 S 巯基转移酶融合表达载体pGEX 4T 2 ,构建成重组表达质粒pGEX TC。经SDS PAGE蛋白电泳鉴定 ,表达产物为 76Kd的特异性重组蛋白。免疫印迹实验证实 ,重组蛋白抗原是破伤风C片段抗原。动物实验证明此重组抗原具有良好的免疫原性 ,1μg免疫剂量可使动物产生 1～ 2IU/ml的破伤风抗毒素 ,ELISA抗体滴度可达 1∶80 0。
The fragment C gene of tetanus toxin was amplified from plasmid DNA of Clostridium tetani by PCR. deteced by DNA sequencing, then cloned into vector pGEX 4T 2 to construct a recombinant plasmid pGEX TC。 SPS PAGE showed that a specific recombinant protein with a molecular weight of 76000 was expressed. The protein antigen was proved to be tetanus toxin fragment C specific antigen by Western blotting. Animal test showed good immunogenicity of the recombinant antigen. The antigen at a dosage of 1 μg could induce 1 2 IU/ml of antitoxin with an ELISA titer of 1∶800.
出处
《中国生物制品学杂志》
CAS
CSCD
1999年第3期133-135,共3页
Chinese Journal of Biologicals
