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人(2'-5')寡腺苷酸合成酶克隆cDNA的结构与表达

Structure and expression of a cloned cDNA for human (2'-5')oligoadenylate synthetase.

作者信息

Shiojiri S, Fukunaga R, Ichii Y, Sokawa Y

出版信息

J Biochem. 1986 May;99(5):1455-64. doi: 10.1093/oxfordjournals.jbchem.a135615.

Abstract

17S poly(A)+RNA, which hybridized to an oligonucleotide complementary to a part of the partial cDNA (E1cDNA) (Merlin et al. (1983) Proc. Natl. Acad. Sci. U.S. 80, 4904) for 2-5A synthetase, was isolated from interferon-treated human KB cells and used for cDNA cloning. Several overlapping cDNAs were cloned by using the oligonucleotide as a probe. Two of them were joined at their overlapping region, resulting in a cDNA (22-1 cDNA) of 1.4 kb containing a long open reading frame. When the cDNA was expressed in COS-7 cells with an eukaryotic promoter, active 2-5A synthetase was produced and localized mainly in the cytoplasm. The 5'-proximal ATG in 22-1 cDNA is followed immediately by another ATG. This second ATG was assumed to work as the initiator codon. If so, this enzyme comprises 363 amino acids.

摘要

从经干扰素处理的人KB细胞中分离出与2-5A合成酶部分cDNA(E1cDNA)(梅林等人,(1983年)《美国国家科学院院刊》80卷,4904页)的一部分互补的寡核苷酸杂交的17S多聚腺苷酸加尾RNA,并将其用于cDNA克隆。使用该寡核苷酸作为探针克隆了几个重叠的cDNA。其中两个在其重叠区域连接,产生了一个1.4 kb的cDNA(22-1 cDNA),其含有一个长的开放阅读框。当该cDNA在具有真核启动子的COS-7细胞中表达时,产生了有活性的2-5A合成酶,并且主要定位于细胞质中。22-1 cDNA中5'近端的ATG紧接着另一个ATG。这个第二个ATG被认为是起始密码子。如果是这样,这种酶由363个氨基酸组成。

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