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与牛催乳素信使核糖核酸互补的DNA的克隆

Cloning of DNA complementary to bovine prolactin mRNA.

作者信息

Miller W L, Thirion J P, Martial J A

出版信息

Endocrinology. 1980 Sep;107(3):851-3. doi: 10.1210/endo-107-3-851.

Abstract

We have cloned DNA complementary to mRNA coding for bovine prolactin (bPrl). Double-stranded cDNA prepared from bovine pituitary mRNA was inserted into the Pst I site of plasmid bPR322 by the dC x dG tailing technique and amplified in E. coli chi 1776. A recombinant plasmid containing bPrl cDNa was identified by hybridization to cloned rat Prl cDNA. It contains cDNA corresponding to the region of the mRNA coding for the carboxy terminal 101 amino acids of bPrl, as well as 42 nucleotides in the 3' untranslated region of the mRNA. Nucleotide sequencing confirmed the amino acid sequencing of this region of bPrl, and permitted the assignment of asparagine or glutamic acid at seven previously equivocal loci. Codon use in bPrl mRNA is comparable to that found in rat and human Prl mRNA's and differs from that in bovine, rat, and human growth hormone mRNA's.

摘要

我们已克隆出与编码牛催乳素(bPrl)的mRNA互补的DNA。从牛垂体mRNA制备的双链cDNA通过dC×dG加尾技术插入质粒bPR322的Pst I位点,并在大肠杆菌chi 1776中扩增。通过与克隆的大鼠催乳素cDNA杂交鉴定出含有bPrl cDNA的重组质粒。它包含与编码bPrl羧基末端101个氨基酸的mRNA区域相对应的cDNA,以及mRNA 3'非翻译区的42个核苷酸。核苷酸测序证实了bPrl该区域的氨基酸序列,并确定了七个先前不确定位点上的天冬酰胺或谷氨酸。bPrl mRNA中的密码子使用情况与大鼠和人类催乳素mRNA中的情况相当,与牛、大鼠和人类生长激素mRNA中的情况不同。

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