Chakrabarti S, Srinivasan J, Lall L, Rao L V, Talwar G P
Gene Expression Laboratory, National Institute of Immunology, New Delhi, India.
Gene. 1989 Apr 15;77(1):87-93. doi: 10.1016/0378-1119(89)90362-4.
Vaccinia virus (VV) expression vector was used to clone the genes for coding alpha and beta subunits of human chorionic gonadotropin (hCG). Recombinant viruses VSL3 and VSS1 containing these genes were selected as blue coloured plaques on the basis of co-expression of Escherichia coli beta-galactosidase in the infected cells. CV-1 cells when infected with VSL3 or VSS1 secreted 2.4 and 1.8 micrograms of alpha and beta hCG subunits, respectively, per 3 x 10(6) cells after 24 h of infection. The subunit proteins expressed individually had immunoreactivity with monoclonal and polyclonal antibodies specific to hCG. The subunit hormonal peptides associated with each other during co-infection to form the complete hCG dimer, which was biologically active as evident from the induction of steroidogenesis in a mouse Leydig cell system.
痘苗病毒(VV)表达载体用于克隆编码人绒毛膜促性腺激素(hCG)α和β亚基的基因。基于感染细胞中共表达大肠杆菌β-半乳糖苷酶,选择含有这些基因的重组病毒VSL3和VSS1作为蓝色噬菌斑。CV-1细胞在感染VSL3或VSS1后,感染24小时后,每3×10⁶个细胞分别分泌2.4微克和1.8微克的α和β hCG亚基。单独表达的亚基蛋白与hCG特异性的单克隆和多克隆抗体具有免疫反应性。在共感染期间,亚基激素肽相互结合形成完整的hCG二聚体,从在小鼠睾丸间质细胞系统中诱导类固醇生成可明显看出其具有生物活性。