摘要
利用聚合酶链反应特异性扩增编码乙型肝炎表面抗原(HBsAg)羧基末端124个氨基酸的基因片段,并将其在大肠杆菌中获得了高效表达。表达产物氨基末端有1个含6组氨酸肽段的20氨基酸融合部分,分子量16178Da,以包含体形式存在,约占细菌总蛋白的22%。以HiTrap亲和层析纯化,纯度可达95%以上。分别用抗HBsAg多克隆抗体和抗HBsAga单克隆抗体进行ELISA检测表明,该蛋白具有HBsAg反应原性。
The gene fragment coding the 124 amino acids of the carboxyl terminus of hepatitis B surface antigen(HBsAg)was amplified by polymerase chain reaction(PCR)and the shorted HBsAg was expressed in E.coli in a fusion protein fashion.The fusion protein was 16178Da in molecular weight and contained a 20 amino acid fusion portion at its amino terminus which contained a six histidine peptide segment.The expressed proteins aggregated into inclusion bodies weigh about 22% of the total bacteria proteins.It was easily purified with the HiTrap affinity matrix with a purity up to 95%.ELISA detection showed that the protein could be recognized by polyclonal antibodies against HBsAg and monoclonal antibody against HBsAg a determinant.
出处
《免疫学杂志》
CAS
CSCD
北大核心
1999年第3期180-182,共3页
Immunological Journal