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人γ-干扰素(IFN-γ)cDNA存在序列多态性
Sequence Polymorphism of Human Interferon-γ cDNA
【摘要】 根据已发现的分子进化规律推测,人IFN-γcDNA序列应存在多态性。为检验此推测,我们进行了中国人IFN-γcDNA的克隆与序列分析。用PHA、TPA协同刺激正常人外周血淋巴细胞,从中提取总RNA,经反转录PCR扩增出不含信号肽的IFN-γcDNA片段,于EcoRI和BamHI位点克隆入pUC 19载体中。双向序列测定表明,中国人IFN-γcDNA与国外报道的序列在三个位点上不同:185位A→G(Lys→Arg),399位A→G(氨基酸未变),422位A→G(Gln→Arg)。此结果证明我们的推测正确。
【Abstract】 According to the principles of molecular evolution, human interferon-γ(IFN-γ) cDNA should have sequence polymorphism. To test this inference, we carried out cDNA cloning and sequencing of Chinese IFN-γ. At first, the peripheral blood lymphocytes of normal volunteers were stimulated synergistically with PHA and TPA. Then, total RNA was isolated from these cells and used for reverse-transcription (RT)-PCR. The amplified cDNA of human mature IFN-γ was digested with EcoRI and BamHI, and cloned into pUC19. Sequencing of individual clones from both direction showed that Chinese IFN-γ cDNA was different from the sequence of other nations at three nucleotides reported previously: No. 185, A→G (Lys→Arg); No. 399, A→G (without change of amino acid); No. 422, A→G (Gln→Arg). The results above verified our inference made from the principles of molecular evolution.
【Key words】 Human interferon-γ; RT-PCR; cDNA cloning; Sequencing; Principles of molecular evolution;
- 【文献出处】 高技术通讯 ,High Technology Letters , 编辑部邮箱 ,1993年11期
- 【分类号】Q75
- 【被引频次】1
- 【下载频次】17