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通过寡核苷酸合成仪,用化学方法合成相当于hcGβ链C末端36肽基因的片段,成功地克隆到噬菌体载体λgt11中,并在宿主菌株E. coli y_(1089)中得到表达。含λgt11 hCG重组体的E. coli y_(1089)所合成的含人类绒毛膜促性腺激素β链c末36肽的杂交蛋白(下称:β—galactosidase—hCG),通过硫酸铵沉淀、亲和层析和制备性聚丙烯酰胺凝胶电泳,可得电泳纯单一色带的β—gaIactosidase—hCG。蛋白质转印(Western Blot)技术证实β—galactasidase—hCG具有hCGβ链c末端36肽的免疫特性。对流免疫电泳和免疫双扩散试验结果表明β—galactosidase—hCG具免疫原性。从一立升培养液的菌体中可分离纯化约3.8毫克的β—galaetosidase—hCG。
A fragment corresponding to the 36 peptide gene of C-terminal of hcGβ chain was chemically synthesized by an oligonucleotide synthesizer and successfully cloned into the bacteriophage vector λgt11 and expressed in the host strain E. coli y_ (1089). The hybrid protein containing human chorionic gonadotropin beta chain c-terminal peptide 36 (hereinafter, referred to as: β-galactosidase-hCG) synthesized by E. coli y_ (1089) containing λgt11 hCG recombinant was purified by ammonium sulfate precipitation, affinity Chromatography and preparative polyacrylamide gel electrophoresis gave β-gaIactosidase-hCG as a single, single-band electrophoresis. Western Blot technology confirmed that β-galactasidase-hCG has the immunogenic properties of the hCGβ chain c-terminal 36 peptide. Convective immunoelectrophoresis and immunodiffusion double diffusion test results show that β-galactosidase-hCG is immunogenic. About 3.8 mg of β-galactosidase-hCG can be isolated and purified from a single liter of culture broth.