Zarucki-Schulz T, Kulomaa M S, Headon D R, Weigel N L, Baez M, Edwards D P, McGuire W L, Schrader W T, O'Malley B W
Proc Natl Acad Sci U S A. 1984 Oct;81(20):6358-62. doi: 10.1073/pnas.81.20.6358.
A cDNA for the chicken progesterone receptor B subunit antigen (Mr, 108,000) has been isolated from a cDNA library prepared from size-selected chicken oviduct poly(A)+RNA. A specific monoclonal antibody raised against hen progesterone receptor B subunit (alpha PR-B) was used to screen the library. Recombinant clones reacting with the antibody by virtue of antigen expression were used in hybrid-selected translation. A single clone, pPRB-1, hybridized specifically to a mRNA that yielded a Mr 108,000 protein when translated in vitro and which was immunoprecipitable by the alpha PR-B antibody. This cDNA represents a 470-base-pair portion of the PR-B nucleotide sequence. Additional clones have been subsequently isolated from the recombinant library using the insert from pPRB-1 as a specific probe. A mRNA size of approximately 3000 nucleotides was determined for the chicken progesterone receptor B subunit by formaldehyde/agarose gel electrophoresis and blot hybridization using pPRB-1 as a probe. Preliminary studies show that withdrawal of hormone from chickens treated chronically with estrogen leads to a dramatic decrease in the cellular RNA concentration of receptor B, indicating that target tissue levels of receptor B RNA are under hormonal control.
已从经大小选择的鸡输卵管多聚腺苷酸加尾RNA制备的cDNA文库中分离出鸡孕酮受体B亚基抗原(分子量108,000)的cDNA。用针对母鸡孕酮受体B亚基(αPR - B)产生的特异性单克隆抗体筛选该文库。通过抗原表达与抗体反应的重组克隆用于杂交选择翻译。单个克隆pPRB - 1与一种mRNA特异性杂交,该mRNA在体外翻译时产生分子量为108,000的蛋白质,并且可被αPR - B抗体免疫沉淀。该cDNA代表PR - B核苷酸序列的470个碱基对部分。随后使用pPRB - 1的插入片段作为特异性探针从重组文库中分离出其他克隆。通过甲醛/琼脂糖凝胶电泳并用pPRB - 1作为探针进行印迹杂交,确定鸡孕酮受体B亚基的mRNA大小约为3000个核苷酸。初步研究表明,长期用雌激素处理的鸡去除激素后,受体B的细胞RNA浓度会急剧下降,这表明受体B RNA的靶组织水平受激素控制。