摘要
基因工程菌株产生的含人类绒毛膜促性腺激素(hCG)β亚单位C末端36肽的β-半乳糖苷酶杂交蛋白(简称β-galaclosidase-hCG),在未经解聚处理前免疫新西兰种白兔和昆明种小白鼠,其抗血清在免疫双扩散试验和对流免疫电泳中对hCG没有形成沉淀线作用。而此杂交蛋白经解聚处理后,其抗血清与hCG相作用,在对流免疫电泳和免疫双扩散试验中都出现抗原抗体反应的免疫沉淀线。从多种蛋白质解聚系统中,经过反复试验,筛选出对β-galactosidase-hCG杂交蛋白有良好解聚作用的解聚剂为2M脲、0.9MSDS、0.005M巯基乙醇;解聚作用最适温度是60℃,最适时间为40分钟。
An artificial gene equivalent to 36 C-terminal peptide of human chorionic gonadotropin (hCG) β-subunit has been chemically synthesized through the application oligonucleotide synthesizer and the double strand DNA has heen successfully cloned into an expression vector, λgtll, and transfected into the host strain E.Coli Y1089, through recombinant phage λgtll hCG to produce a hybrid protein containing hCG-β-suhunit 36 C-terminal peptide (Called β-galactosidase-hCG). Anti-serum of rat immunized with β-galactosidase-hCG was detected by counter current electrophoresis and immunodiffusion, the results hare clearly demonstrated that β-galactosidase hCG can be crossly immunoprecipitated with rat anti-β-galactosidase-hCG serum, whereas using hCG replaced β-galactosidase-hCG, the anti-serum produced against β-galactosidase-hCG has no cross reaction with hCG. When β-galactosidase-hCG was treated with 2M Urea, 0.9M SDS and 0.005M Mercaptoethanol solvent system before using as an antigen to immunize rat, the anti-body against this depolymerized antigen can be crossly immunoprecipitated with hCG. It revealed that the β-galactosidase-hCG polymer was depolymerized and the conformation of β-galactosidase-hCG subunit has been changed, therefore the antigenic determinant of hCG-β-subunit 36C-terminal peptide in β-galactosidase-hCG appeared its immunogenicity in Urea and SDS solvent. the depolymerized optimal condictions for, β-galaztosidase-hCG in Urea and SDS solvent system were 60C in 40 minutes.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
1989年第1期53-59,共7页
Journal of Jinan University(Natural Science & Medicine Edition)
关键词
HCG
B-半乳糖苷酶
SDS免疫原性
β-hCG C-terminal peptide β-Galactosdase, Urea, Dodeeyl sodium sulfate, Mereaptcethanol, Immunogenicity
