Hela细胞P16^(ink4)cDNA的克隆及序列分析

Molecular Cloning and Sequencing of P16^(ink4) cDNA from Hela Cell

P16^(ink4)是CDK抑制蛋白,参与调控细胞G1期至S期的转换。目前发现P16`(ink4)基因损伤与多种肿瘤的发生、发展有关,可能是功能上非常重要的抑癌基因。为了研究该基因的功能,以及突变对该基因功能的影响,本文应用RT-PCR方法,从Hela细胞中克隆了P16^(ink4)cDNA。扩增得到556bp片段(包括引物两端酶切位点的16bp)克隆于M13载体,测定了其DNA序列。该序列包括了P16^(ink4)cDNA编码区全部471bp,以及3’端69bp序列。表明P16^(ink4)cDNA已成功地得到克隆。

P16ink4 is one of the CDK (cyclin dependent kinase) inhabitor. which inhabits theability of CDK to interact with cyclin D and stimulate passage through the G1 phase of the cell cycle. Recently, the observation of a very high frequency of P16'nk4 gene mutations in many tumor type ssuggest that the gene is a strong candidate for tumor suppressor gene. For studing the function of the P16ink4 gene, we cloned P16ink4 cDNA from HeLa cell by RT-PCR method. The P16ink4 cDNA which cloned from HeLa cell includes 471 bp open reading frame and 3' 69 bp non coding region. The P16ink4 cDNA Sequence is indentical to the published sequence.