Molecular cloning and nucleotide sequence of full-length of cDNA coding for porcine gastrin.
Molecular cloning and nucleotide sequence of full-length of cDNA coding for porcine gastrin.
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猪胃泌素编码cDNA全长的分子克隆和核苷酸序列。
DOI：
10.1073/pnas.79.4.1049
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发表时间：
1982
影响因子：
11.1
通讯作者：
Agarwal,KL
中科院分区：
文献类型：
--
作者：
Yoo,OJ;Powell,CT;Agarwal,KL
我们在大肠杆菌中克隆了一个编码猪胃窦黏膜激素前泌素的mRNA cDNA拷贝。合成了全长双链cDNA，利用均聚延伸技术将其插入质粒pBR322的Pst I核酸内切酶位点。以部分cDNA克隆为探针，通过集落杂交在cDNA文库中鉴定完整cDNA克隆。分离到4个阳性克隆，其中1个与猪前胃泌素mRNA对应。cDNA插入片段（602个核苷酸）的核苷酸序列显示，整个mRNA编码区有312个核苷酸，5‘非翻译区有61个核苷酸，3’非翻译区有86个核苷酸，poly(a) tail有86个核苷酸。胃泌素位于前胃泌素的羧基端附近，在其氨基端和羧基端两侧都有一对碱性氨基酸残基。胃泌素的苯丙氨酸羧基端邻有甘氨酸和一对碱基氨基酸残基，表明苯丙氨酸酶酰胺化反应可能需要甘氨酸和一对碱基氨基酸残基。
We have cloned in EScherichia coli a cDNA copy of mRNA coding for the porcine antral mucosal hormone preprogastrin. Full-length double-stranded cDNA was synthesized and inserted into the Pst I endonuclease site in plasmid pBR322 by using a homopolymeric extension technique. A partial cDNA clone was used as a probe to identify a complete cDNA clone in a cDNA library by colony hybridization. Four positive clones were isolated, one of which corresponded to porcine preprogastrin mRNA. The nucleotide sequence of the cDNA insert (602 nucleotides) revealed 312 nucleotides in the entire mRNA coding region, 61 nucleotides in the 5' untranslated region, 86 nucleotides in the 3'untranslated region, and a poly(A) tail of 86 nucleotides. Gastrin is located near the carboxyl end of preprogastrin and is flanked at both its amino and carboxyl ends by a pair of basic amino acid residues. The presence of glycine and a pair of basic amino acid residues adjacent in the carboxyl-terminal phenylalanine of gastrin indicates that the glycine and a pair of basic amino acid residues may be required for the enzymatic amidation of phenylalanine to phenylalanine amide.