摘要
将 PCR扩增得到的 pc MV- S中的乙肝表面抗原 (HBs Ag) S基因亚克隆到 p UC18中 ,构建成 p U S质粒 ,再将化学合成的生长抑素 (somatostatin,SS)基因单链复性 ,融合到 S基因编码区的第 2 2 5个氨基酸位点之后 ,构建成 p US/SS质粒 ,然后将 S/ SS融合基因亚克隆到 pc DNA3.1(- )中 ,构建成 S/ SS融合表达质粒 pc S/ SS。采用脂质体包裹法将 pc S/ SS质粒转染 He L a细胞 ,用 SDS- PAGE和 EL ISA分析表明 ,S/ SS融合基因在转染后 72 h和 G4 18选择 2周后的细胞中均获得表达 ,融合蛋白具有 SS的反应原性。
The S gene of HBsAg in pcMV-S was amplified by PCR and subcloned into pUC18,and the resultant recombinant plasmid was pUS.The single strands of somatostatin(SS) gene synthesized chemically were annealed and fused to the downstream of 225 amino acids of the encoding region of S gene and then plasmid pUS/SS was constructed.Subsequently the fused gene of S/SS was subcloned into pcDNA3.1(-),an eukaryotic expression vector,and the resultant plasmid pcS/SS was used to transfect HeLa cells through liposome embedding.SDS-PAGE and ELISA results revealed S/SS fused gene expressed transiently in cells at 72 h after transfection,and the fused protein which possessed the immunological activity of somatostatin was also detected in transfected cells after two weeks of G418 selection.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2004年第2期153-156,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目 (3 0 2 70 95 9)
