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利用恒定区的聚合酶链反应片段对猪T细胞受体α、β、γ和δ链进行分子克隆。

Molecular cloning of porcine T cell receptor alpha, beta, gamma and delta chains using polymerase chain reaction fragments of the constant regions.

作者信息

Thome A, Saalmüller A, Pfaff E

机构信息

Federal Research Center for Virus Diseases of Animals, Tübingen, FRG.

出版信息

Eur J Immunol. 1993 May;23(5):1005-10. doi: 10.1002/eji.1830230503.

Abstract

Fragments of the constant regions of porcine alpha, beta, delta and two types of gamma T cell receptor (TcR) chains were obtained by reverse transcription and polymerase chain reaction (PCR). Screening of a porcine peripheral T cell cDNA library with these PCR fragments led to the isolation of porcine TcR alpha, beta, gamma and delta chain clones. Sequence analysis of these clones and the respective PCR fragments demonstrated the existence of one alpha, one beta, three gamma and one delta chain isotype. Comparisons of the deduced amino acid sequences of the constant region with other species revealed a significant homology. For two of the three identified porcine gamma chain insertions of 38 and 40 amino acids were found within the hinge region. In addition, our sequence data demonstrate a high variability in the cytoplasmic C gamma domain among the three porcine TcR gamma isotypes as well as between species, which might be of structural and/or functional significance. Comparison with biochemical data indicate the existence of four porcine TcR gamma isotypes.

摘要

通过逆转录和聚合酶链反应(PCR)获得了猪α、β、δ和两种类型γ T细胞受体(TcR)链恒定区的片段。用这些PCR片段筛选猪外周血T细胞cDNA文库,从而分离出猪TcRα、β、γ和δ链克隆。对这些克隆和相应的PCR片段进行序列分析,结果表明存在一种α链、一种β链、三种γ链和一种δ链同种型。将恒定区推导的氨基酸序列与其他物种进行比较,发现存在显著的同源性。在已鉴定的三条猪γ链中,有两条在铰链区内发现了38和40个氨基酸的插入。此外,我们的序列数据表明,三种猪TcRγ同种型之间以及不同物种之间,胞质Cγ结构域具有高度变异性,这可能具有结构和/或功能意义。与生化数据比较表明存在四种猪TcRγ同种型。

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