应用抑制性消减杂交技术克隆人肺鳞癌差异性表达基因(附件).PDFVIP

·10 · 中国肺癌杂志2002 年 2 月第 5 卷第 1 期　Chin J Lung Cancer , February 2002 ,Vol. 5 , No. 1 ·基础研究· 应用抑制性消减杂交技术克隆人肺鳞癌 差异性表达基因 沈晨阳 　刘军 　王丹蕾 　赵辉 　姜冠潮 　王俊 　张国良 ( 【摘要】　目的 　克隆人肺鳞癌肿瘤差异性表达基因。方法 　应用抑制性消减杂交技术 suppression sub ) tractive hybridization ,SSH 构建人肺鳞癌 cDNA 消减杂交文库 ,筛选后挑选阳性克隆进行测序 ,并在 Gen Bank 数 ( 据库中进行同源性比较 ,最后经 RTPCR 验证。结果 　成功克隆 10 个差异基因片段 ,其中 2 个为新基因 Gen ) Bank 登录号分别为 :C20 :AF363068 ;C34 :AY032661 。结论 　抑制性消减杂交技术是克隆差异表达基因及发现 新基因的有效方法。 【关键词】　抑制性消减杂交技术 　　肺鳞状细胞癌 　　克隆 　　互补 DNA 【中图分类号】　R734. 2 ;R7335 + 4 Molecular cloning of genes differentially expressed in human lung squamous cell carcinoma using suppression sub tractive hybridization 　SHEN Chenyang , LIU Jun , WANGDanlei , ZHAO Hui , JIANG Guanchao , WANGJun , ZHANG Guoliang. Department of Thoracic Surgery , The People ’s Hospital , Peking University , Beijing 100044 , P . R . China 【Abstract 】　Objective 　To clone and identify genes differentially expressed in human lung squamous cell carcinoma (LSCC) . Methods 　A subtracted cDNA library of human LSCC was constructed by suppression subtracted hybridization ( SSH) method. After screening , the subtracted library clones representing mRNAs that were truly differentially expressed in LSCC but not in its adjacent noncancerous tissues were selected to identify by RTPCR and DNA sequencing were per formed. Nucleic acid homology searches were performed using the BLAST program. Results 　By this technique , 10 differ entially expressed gene cDNA fragments of LSSC were obtained. Two were novel and eight were al