应用混合探针文库筛选法克隆多个肿瘤差异表达基因  

Isolation of tumor differentially expressed genes by mixing probes library screen

作　　者：余鹰[1] 谢奕[1] 朱诗国 周鸣[1] 张必成 李桂源[1] public.cs.hn.cn 

机构地区：[1]湖南医科大学肿瘤研究所,湖南长沙410078

出　　处：《癌症》2000年第7期709-712,共4页Chinese Journal of Cancer

基　　金：国家863项目!102-10-01-05;973重点项目!<"疾病基因组学"理论和技术体系的建立>;自然科学基金!39700158

摘　　要：目的：进一步分离在鼻咽癌组织中表达下调／缺失的候选抑瘤基因。方法：采用PCR方法对有差异表达的cDNA序列（AF152605和AF091517）进行探针标记，Northern杂交确定其mRNA转录本大小，进而混合两种PCR探针对骨骼肌cDNA文库进行筛选，阳性克隆经PCR鉴定后直接测序。结果：克隆了转录本为2．2Kb、1．1Kb和1．4Kb三个基因，GenBank登录号分别为AF179285、AF170307和AF194971。结论：混合探针文库筛选法是同时、快速获得多个cDNA片段其全长序列的可借鉴实验手段。Objective: To clone candidate tumor suppressor genes down-expressed in nasopheqngeal carcinoma (NPC). Methods: Differentially expressed cDNA fragments (AF152605 and AF091517) were labeled by PCR, and Northern blot was used to coallrm transcript length of these genes. Skeleton muscle cDNA library was screened through PCR-labeled probe mixture. Results: 23 indePendently positive and overiapping positive clones were obtained. By sequencing the positive clones directly, three novel genes (Genbank accession number: AF179285, AF 170307 and AF194971 ), with transcription sizes of 2. 1 Kb, 1. 1Kb and 1. 4 Kb respectively, were isolated successfully.Conclusions: Library screen using PCR-labeled probe mixture is an available method to get full-length cDNA from multi-cDNA fragment simultaneously and quickly.

关 键 词：鼻咽肿瘤 文库筛选 抑瘤基因 基因克隆 

分 类 号：R730.3[医药卫生—肿瘤]