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牛催乳素信使核糖核酸的核苷酸序列。序列多态性的证据。

Nucleotide sequence of bovine prolactin messenger RNA. Evidence for sequence polymorphism.

作者信息

Sasavage N L, Nilson J H, Horowitz S, Rottman F M

出版信息

J Biol Chem. 1982 Jan 25;257(2):678-81.

PMID:6274859
Abstract

Hybrid molecules containing DNA sequences complementary to bovine pituitary mRNA were constructed in the Pst I site of pBR322 by the dC . dG tailing technique. Recombinant plasmids containing bovine prolactin (bPRL) sequences were amplified in bacteria and identified by hybridization to purified [32P]bPRL cDNA sequences. Nucleotide sequence analysis was performed on the inserts from two of the positive clones. One clone, pBPRL72, contained a 982-base pair insert that included 67 nucleotides of the 5'-untranslated region, the complete coding region of the preprolactin protein (690 nucleotides), and the entire 3'-untranslated region (150 nucleotides) of bPRL mRNA. The nucleotide sequence analysis of clone pBPRL72 predicted the sequence of a 30-amino acid signal peptide and confirmed the published amino acid sequence of the protein with one exception. A comparison of the pBPRL72 cDNA sequence with a second bPRL clone, pBPRL4, revealed four silent nucleotide differences. Three of the base changes occurred in the third position of amino acid codons, and one occurred in the 3'-noncoding region. The sequence polymorphism suggests the existence of alleles or multiple loci for bPRL that do not alter the protein structure.

摘要

利用dC·dG加尾技术,在pBR322的Pst I位点构建了含有与牛垂体mRNA互补DNA序列的杂合分子。含有牛催乳素(bPRL)序列的重组质粒在细菌中扩增,并通过与纯化的[32P]bPRL cDNA序列杂交进行鉴定。对两个阳性克隆的插入片段进行了核苷酸序列分析。一个克隆pBPRL72含有一个982个碱基对的插入片段,其中包括5'-非翻译区的67个核苷酸、前催乳素蛋白的完整编码区(690个核苷酸)以及bPRL mRNA的整个3'-非翻译区(150个核苷酸)。克隆pBPRL72的核苷酸序列分析预测了一个30个氨基酸的信号肽序列,并证实了已发表的该蛋白氨基酸序列,只有一处例外。将pBPRL72 cDNA序列与第二个bPRL克隆pBPRL4进行比较,发现了四个沉默核苷酸差异。其中三个碱基变化发生在氨基酸密码子的第三位,一个发生在3'-非编码区。序列多态性表明存在不改变蛋白质结构的bPRL等位基因或多个基因座。

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