摘要
本研究根据鸡CD4分子胞外区基因序列设计合成一对引物,用PCR技术从三黄鸡的cDNA文库中克隆出了其CD4胞外区基因片段,大小为1 206 bp,将该片段插入pMAL-p2X表达载体中,转化大肠杆菌TB1感受态细胞后进行诱导表达,获得了分子量约为87.5 kD的融合蛋白。表达产物经Amylose树脂亲和层析柱纯化,得到了高纯度的鸡CD4胞外区的融合蛋白。利用此蛋白免疫小鼠,制备了高效价的鼠源抗鸡CD4胞外区的抗体,结果表明该蛋白具有良好的抗原性。
The gene fragment of chicken CD4 extracellular region was Cloned from cDNA library using a pair of primers. It was 1206 bp in length and inserted into pMAL-p2X prokaryotic system to form a recombinant plasmid. The recombinant plasmid was transformed into E. coli TB1 and the fusion protein was expressed by induction. The molecular weight（MW） of this fusion protein was 87.5 kD as analyzed by SDS-PAGE and the purified fusion protein,which was purified by amylose affinity resin column prior to immunizing mice and the specific antibodies were produced, measured by ELISA. The results show that the fusion protein was of antigenicity.
出处
《中国兽医杂志》
CAS
北大核心
2007年第4期3-5,共3页
Chinese Journal of Veterinary Medicine
关键词
鸡CD4
胞外区
克隆
原核表达
chicken
CD4
extracellular region
clone
prokaryotic expression
