用限制性cDNA文库制作K562细胞基因表达谱芯片探针  

Probes-Preparation of cDNA Microarray for the Gene Expression in Leukemia K562 Cell using Restriction cDNA Library

作　　者：祝骥[1] 马文丽[1] 李凌 宋艳斌[1] 吴清华[1] 姚汝华[2] 郑文岭[3] 

机构地区：[1]第一军医大学分子生物学研究所,中国广东广州510515 [2]华南理工大学生物工程系,中国广东广州510640 [3]广州军区广州总医院分子肿瘤研究所,中国广东广州510010

出　　处：《生命科学研究》2002年第2期142-146,共5页Life Science Research

基　　金：国家自然科学基金 ( 39880 0 32 );广州市重点科技攻关项目 ( 990 4480 2 2 )资助

摘　　要：以人红白血病K5 6 2细胞为材料 ,应用限制性显示PCR(RD -PCR)技术构建cDNA文库 .该文库通过PCR引物 3′端延伸两个不同碱基形成 136对引物对cDNA进行限制性扩增 ,得到 136组不同的PCR扩增产物 ,纯化后与载体连接并转化细菌 ,即为限制性cDNA文库 .根据不同的分组进行克隆的鉴定和分离 ,并进行大量扩增制备cDNA芯片探针 .该方法构建的文库因经过了限制性分组扩增 ,每组均含有特定的cDNA ,因而大大加快了随后克隆的分离和鉴定的速度 ,为基因芯片探针制备提供了一个新方法 .Using human leukemia K562 cells, restriction display PCR technology was applied to construct a cDNA library. cDNA was restrictedly amplified by PCR which primers 3′end was extended two different bases to become 136 pair of primers. The PCR products of 136 groups were purified and ligated with T-vector to transform E.coli JM 109 to build up a cDNA library. The clones were identified and isolated according to the different groups, then prepared probes of cDNA microarray by large amplification. Each groups of this library which was amplified by restriction grouping contained specific cDNA, so the identification and isolation of clones were largedly accelerated. That provided a new method for probe-preparation of cDNA microarray.

关 键 词：限制性cDNA文库 K562细胞 基因表达谱芯片 癌变细胞 

分 类 号：Q786[生物学—分子生物学] R394.8[医药卫生—医学遗传学]