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来自中国仓鼠的一个扩增型谷氨酰胺合成酶基因的cDNA克隆及核苷酸序列

The cloning and nucleotide sequence of cDNA for an amplified glutamine synthetase gene from the Chinese hamster.

作者信息

Hayward B E, Hussain A, Wilson R H, Lyons A, Woodcock V, McIntosh B, Harris T J

出版信息

Nucleic Acids Res. 1986 Jan 24;14(2):999-1008. doi: 10.1093/nar/14.2.999.

Abstract

The nucleotide sequence for a glutamine synthetase (GS) mRNA from gene-amplified Chinese hamster (CHO) cells was determined from recombinant cDNA clones obtained from both pBR322 and lambda gt10 libraries and by primer extension. The sequence obtained contains about 1400 bp corresponding to a minor species of mRNA terminated by a poly A sequence. The mRNA contains 146 nucleotides of 5'-noncoding region, 1119 bp of coding sequence, and 108 bp of 3'-noncoding sequence with a 32 bp poly(A) tail. The polyadenylation site used shows little homology with efficient polyadenylation sites, but has considerable complementarity with U4 RNA. The predicted amino acid sequence, starting from an initiation codon with the preferred sequence surrounding it, indicates that Chinese hamster GS has high homology with published bovine brain GS peptides and enabled an ordering of these peptides. There is homology between the mammalian GS enzymes and glutamine synthetases obtained from plants and cyanobacteria but no obvious homology between the CHO cell GS sequence and that of other ATP hydrolysing enzymes.

摘要

通过从pBR322和λgt10文库获得的重组cDNA克隆以及引物延伸法,确定了基因扩增的中国仓鼠(CHO)细胞中谷氨酰胺合成酶(GS)mRNA的核苷酸序列。获得的序列包含约1400个碱基对,对应于一种由聚腺苷酸序列终止的次要mRNA种类。该mRNA包含146个核苷酸的5'-非编码区、1119个碱基对的编码序列以及108个碱基对的3'-非编码序列,带有32个碱基对的聚腺苷酸尾巴。所使用的聚腺苷酸化位点与高效聚腺苷酸化位点几乎没有同源性,但与U4 RNA具有相当的互补性。从具有其周围优选序列的起始密码子开始预测的氨基酸序列表明,中国仓鼠GS与已发表的牛脑GS肽具有高度同源性,并能够对这些肽进行排序。哺乳动物的GS酶与从植物和蓝细菌获得的谷氨酰胺合成酶之间存在同源性,但CHO细胞GS序列与其他ATP水解酶的序列之间没有明显的同源性。

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