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利用cDNA表达载体分离小鼠白细胞介素2 cDNA克隆:猴细胞转染后T细胞生长因子活性的表达

Use of a cDNA expression vector for isolation of mouse interleukin 2 cDNA clones: expression of T-cell growth-factor activity after transfection of monkey cells.

作者信息

Yokota T, Arai N, Lee F, Rennick D, Mosmann T, Arai K

出版信息

Proc Natl Acad Sci U S A. 1985 Jan;82(1):68-72. doi: 10.1073/pnas.82.1.68.

Abstract

A cDNA sequence coding for mouse interleukin 2 (IL-2) has been cloned from a cDNA library prepared from mRNA derived from a concanavalin A-activated mouse T-cell clone. The library was constructed by using the pcD vector system, which permits the expression of cDNA inserts in mammalian cells. Screening of the library was performed by transfecting COS-7 monkey cells with pools of cDNA clones in order to express the products encoded by full-length cDNA inserts. By assaying the supernatant fluid, IL-2 cDNA clones that express T-cell growth-factor (TCGF) activity were identified. The DNA sequence codes for a polypeptide of 169 amino acid residues including a putative signal peptide. The mouse IL-2 amino acid sequence deduced from the nucleotide sequence of its cDNA shares extensive homology with the human IL-2 amino acid sequence reported previously. These results demonstrate that identification of full-length cDNA clones for many lymphokines may be achieved entirely on the basis of detection of the functional polypeptides in mammalian cells.

摘要

已从用伴刀豆球蛋白A激活的小鼠T细胞克隆的mRNA制备的cDNA文库中克隆出编码小鼠白细胞介素2(IL-2)的cDNA序列。该文库是使用pcD载体系统构建的,该系统允许cDNA插入片段在哺乳动物细胞中表达。通过用cDNA克隆池转染COS-7猴细胞来筛选文库,以便表达全长cDNA插入片段编码的产物。通过检测上清液,鉴定出表达T细胞生长因子(TCGF)活性的IL-2 cDNA克隆。该DNA序列编码一个169个氨基酸残基的多肽,包括一个推定的信号肽。从小鼠IL-2 cDNA的核苷酸序列推导的氨基酸序列与先前报道的人IL-2氨基酸序列具有广泛的同源性。这些结果表明,许多淋巴因子的全长cDNA克隆的鉴定可以完全基于在哺乳动物细胞中检测功能性多肽来实现。

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