从经雌二醇处理的雏鸡肝脏中免疫化学分离和鉴定卵黄蛋白原信使核糖核酸

Immunochemical isolation and characterization of vitellogenin mRNA from liver of estradiol-treated chicks.

作者信息

Jost J P, Pehling G

出版信息

Eur J Biochem. 1976 Jul 1;66(2):339-46. doi: 10.1111/j.1432-1033.1976.tb10523.x.

DOI:10.1111/j.1432-1033.1976.tb10523.x

Abstract

Vitellogenin mRNA was purified through three steps. A heavy polysome fraction was obtained by discontinuous sucrose density gradient centrifugation, vitellogenin polysomes were immunoprecipitated with affinity-chromatography-purified anti-lipovitellin IgG and goat anti-rabbit IgG, the enriched mRNA was isolated on a poly(U)-Sepharose column. As judged by its specific activity in a reticulocyte lysate system, vitellogenin mRNA has been enriched a 1000-fold with a recovery of 30%. On 99% formamide 3.4% polyacrylamide gels vitellogenin mRNA has an Mr of 2.4-2.5 X 10(6) and codes for a peptide of Mr 240000, which under our incubation conditions is partially degraded to smaller peptides.

摘要

卵黄原蛋白信使核糖核酸（Vitellogenin mRNA）通过三个步骤进行纯化。通过不连续蔗糖密度梯度离心获得重多核糖体组分，用亲和层析纯化的抗脂卵黄磷蛋白免疫球蛋白G（anti-lipovitellin IgG）和山羊抗兔免疫球蛋白G免疫沉淀卵黄原蛋白多核糖体，在聚尿苷酸-琼脂糖（poly(U)-Sepharose）柱上分离富集的信使核糖核酸。根据其在网织红细胞裂解物系统中的比活性判断，卵黄原蛋白信使核糖核酸已富集1000倍，回收率为30%。在99%甲酰胺、3.4%聚丙烯酰胺凝胶上，卵黄原蛋白信使核糖核酸的分子量为2.4 - 2.5×10⁶，编码一个分子量为240000的肽，在我们的孵育条件下，该肽会部分降解为较小的肽。