Enmark E, Kainu T, Pelto-Huikko M, Gustafsson J A
Center for Biotechnology, Karolinska Institute, Novum, Huddinge, Sweden.
Biochem Biophys Res Commun. 1994 Oct 14;204(1):49-56. doi: 10.1006/bbrc.1994.2424.
Degenerate PCR primers were designed based upon conserved regions within the DNA and ligand binding domains of several nuclear receptors. PCR was performed using these primers starting with total brain cDNA. Several members of the nuclear receptor superfamily were identified, one of which was novel, but showing a high degree of homology to the previously known orphan receptor Rev-ErbA. This new orphan receptor, Rev-ErbA beta, was further characterised, and a cDNA with an intact open reading frame was isolated from a rat brain cDNA library. Northern blot analysis shows two different sizes of mRNA, 4 and 5.5 kb, respectively, in all tissues analysed. A more detailed study of this orphan was then performed using in situ hybridisation, showing a high expression of the new orphan receptor in particular in the cerebellum, the dentate gyrus of the hippocampus and pituitary gland of adult rats.
基于几种核受体的DNA和配体结合域内的保守区域设计了简并PCR引物。使用这些引物从全脑cDNA开始进行PCR。鉴定出了核受体超家族的几个成员,其中一个是新的,但与先前已知的孤儿受体Rev-ErbA具有高度同源性。对这个新的孤儿受体Rev-ErbAβ进行了进一步表征,并从大鼠脑cDNA文库中分离出了一个具有完整开放阅读框的cDNA。Northern印迹分析显示,在所有分析的组织中分别有4 kb和5.5 kb两种不同大小的mRNA。然后使用原位杂交对这个孤儿受体进行了更详细的研究,结果表明该新孤儿受体在成年大鼠的小脑、海马齿状回和垂体中高表达。