人K562细胞cDNA文库的扩增、纯化、鉴定和酵母细胞转化.pdfVIP

重庆医科大学学报2008年第33卷第l期 (Journal of Chongqing Medical University 2008．Vo1．33 No．1) 一 13一 论 著 文章编号：0253—3626(2008)01—0013—05 人K562细胞cDNA文库的扩增、纯化、鉴定和酵母细胞转化 欧阳峰·-一，王东生·，郝 坡·，刘北忠·，刘 畅-，钟 梁·，金丹婷·，王 种· (1．重庆医科大学医学检验系省部共建教育部重点实验室，重庆 400016；2．广东省人民医院皮肤科实验室，广州 510600) 【摘 要】目的：对预转化到大肠杆菌DH5 中的人K562细胞cDNA文库进行扩增、纯化和鉴定并将文库质粒转化酵母细胞， 为下一步的靶蛋白筛选做准备。方法：对K562细胞cDNA文库进行扩增，提取文库质粒，将文库质粒转化酵母Y187细胞，并用 PCR和酶切鉴定转化结果。结果：成功的扩增人K562细胞cDNA文库、并验证了文库的多样性、将文库质粒成功转化酵母 Y187细胞。结论：K562细胞cDNA文库的成功扩增、纯化、鉴定，为进一步的文库筛选奠定了基础。 【关键词】白血病；K562细胞；cDNA文库；多样性 【中国图书分类法分类号】R733．7 【文献标识码】A 【收稿日期】2007—08—29 Amplification，purification and identification of human K562 celI cDNA library and its transformation into yeast cell OUYANG ，et (Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education，Faculty of Laboratory Medicine，Chongqing Medical University) 【Abstract】Objective：To Amplify，purify and identify the cDNA library of human K562 cell in E．coli DH5 and to transform it into yeast cell for subsequent target protein screening．Methods：The cDNA library of human K562 cell Was amplified．Then the library plasmids were extracted and transformed into Y187 yeast cel1．Th e results of transform ation were verified by PCR and restriction enzyme analysis．Results：The cDNA library of human K562 cell Was amplified successfully and the diversities were verified．The cDNA library of human K562 cell was also successfully transformed into Y 1 87 yeast cel1．Conclusion： Successfully amplification，purification and identification of the cDNA library of human K562 cell lay the foundation for the subsequent protein screening． 【Key words】Leukemia；K562 cell line；cDNA library；Diversity 多年来白血病的治疗一直以化学疗法为主，但 胞白血病细胞系进行研究。ph染色体t(9；22)(q34； 化疗的效果有限，相当一部分病例在诱导缓解或巩 ql1)几乎存在于所有的慢粒细胞中。9q34的c—abl 固治疗中发生多药耐药，从而对化疗反应差，易复