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编码大鼠促黄体生成素β亚基前体的cDNA的核苷酸序列。

Nucleotide sequence of the cDNA encoding the precursor of the beta subunit of rat lutropin.

作者信息

Chin W W, Godine J E, Klein D R, Chang A S, Tan L K, Habener J F

出版信息

Proc Natl Acad Sci U S A. 1983 Aug;80(15):4649-53. doi: 10.1073/pnas.80.15.4649.

Abstract

We have determined the nucleotide sequences of cDNAs encoding the precursor of the beta subunit of rat lutropin, a polypeptide hormone that regulates gonadal function, including the development of gametes and the production of steroid sex hormones. The cDNAs were prepared from poly(A)+ RNA derived from the pituitary glands of rats 4 weeks after ovariectomy and were cloned in bacterial plasmids. Bacterial colonies containing transfected plasmids were screened by hybridization with a 32P-labeled cDNA encoding the beta subunit of human chorionic gonadotropin, a protein that is related in structure to lutropin. Several recombinant plasmids were detected that by nucleotide sequence analyses contained coding sequences for the precursor of the beta subunit of lutropin. Complete determination of the nucleotide sequences of these cDNAs, as well as of cDNA reverse-transcribed from pituitary poly(A)+ RNA by using a synthetic pentadecanucleotide as a primer of RNA, provided the entire 141-codon sequence of the precursor of the beta subunit of rat lutropin. The precursor consists of a 20 amino acid leader (signal) peptide and an apoprotein of 121 amino acids. The amino acid sequence of the rat lutropin beta subunit shows similarity to the beta subunits of the ovine/bovine, porcine, and human lutropins (81, 86, and 74% of amino acids identical, respectively). Blot hybridization of pituitary RNAs separated by electrophoresis on agarose gels showed that the mRNA encoding the lutropin beta subunit consists of approximately 700 bases. The availability of cDNAs for both the alpha and beta subunits of lutropin will facilitate studies of the regulation of lutropin expression.

摘要

我们已经测定了编码大鼠促黄体激素β亚基前体的cDNA的核苷酸序列。促黄体激素是一种调节性腺功能的多肽激素,包括配子发育和类固醇性激素的产生。这些cDNA是从卵巢切除术后4周大鼠垂体的聚腺苷酸加尾RNA(poly(A)+RNA)制备而来,并克隆到细菌质粒中。通过与编码人绒毛膜促性腺激素β亚基的32P标记cDNA杂交,筛选含有转染质粒的细菌菌落。人绒毛膜促性腺激素β亚基在结构上与促黄体激素相关。通过核苷酸序列分析检测到几个重组质粒,它们含有促黄体激素β亚基前体的编码序列。对这些cDNA以及使用合成十五聚核苷酸作为RNA引物从垂体聚腺苷酸加尾RNA反转录得到的cDNA的核苷酸序列进行完整测定,得到了大鼠促黄体激素β亚基前体的完整141密码子序列。该前体由一个20个氨基酸的前导(信号)肽和一个121个氨基酸的脱辅基蛋白组成。大鼠促黄体激素β亚基的氨基酸序列与绵羊/牛、猪和人促黄体激素的β亚基相似(分别有81%、86%和74%的氨基酸相同)。在琼脂糖凝胶上电泳分离的垂体RNA的印迹杂交表明,编码促黄体激素β亚基的mRNA约由700个碱基组成。促黄体激素α亚基和β亚基cDNA的可得性将有助于促黄体激素表达调控的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c565/384101/e6d8d794bfcd/pnas00641-0060-a.jpg

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