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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.

机译:cDNA表达载体在分离小鼠白介素2 cDNA克隆中的应用:转染猴细胞后表达T细胞生长因子活性。

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摘要

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.
机译:已经从由伴刀豆球蛋白A活化的小鼠T细胞克隆衍生的mRNA制备的cDNA文库中克隆了编码小鼠白介素2(IL-2)的cDNA序列。该文库是使用pcD载体系统构建的,该系统可在哺乳动物细胞中表达cDNA插入片段。通过用cDNA克隆库转染COS-7猴细胞来进行文库的筛选,以表达全长cDNA插入片段编码的产物。通过分析上清液,鉴定了表达T细胞生长因子(TCGF)活性的IL-2 cDNA克隆。 DNA序列编码包括推定的信号肽的169个氨基酸残基的多肽。从其cDNA的核苷酸序列推导的小鼠IL-2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,对于许多淋巴因子的全长cDNA克隆的鉴定可以完全基于对哺乳动物细胞中功能多肽的检测来实现。

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