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为克隆 7q32 ter区域等位基因杂合性丢失最小共同缺失区的鼻咽癌相关基因 .以STSD7S5 0 9为探针 ,PCR法筛选位于该区的细菌人工染色体 (BAC)克隆 ,应用EST介导的定位 候选克隆策略并结合生物信息学筛选出在鼻咽癌细胞株和活检组织中表达增强的ESTAA7734 5 4,cDNA克隆测序和生物信息学资源获取全长cDNA ,DNA印迹和甲基化分析研究其表达增强的机制 .结果表明 ,克隆的NAG18基因cDNA全长 80 2bp ,编码 2 2 7个氨基酸 ,定位于胞核 .该基因分别与人、鼠TAXREB10 7基因及RPL6基因高度同源 .其表达增强的机制不是基因拷贝数的丢失和甲基化位点的改变 .可以断定NAG18是定位于 7q32 ter最小共同缺失区的鼻咽癌相关基因 ,它是一高度保守的基因 ,参与了DNA的转录活化
To clone cloned nasopharyngeal carcinoma (NPC) genes with minimal loss of heterozygosity in the allele of 7q32 ter region, and to screen for bacterial artificial chromosome (BAC) clones by PCR using STSD7S5 0 9 as a probe and EST mediated Localization of candidate cloning strategies and bioinformatics screening of ESTAA773454 enhanced expression in nasopharyngeal carcinoma cell lines and biopsies 5, cDNA cloning sequencing and bioinformatics resources to obtain full-length cDNA, Southern blot analysis and methylation analysis The results showed that the full-length cDNA of cloned NAG18 gene was 80 2bp, encoding 277 amino acids and located in the nucleus.The gene was highly homologous to human TAXREB107 gene and RPL6 gene, respectively The mechanism of enhancement is not the loss of the gene copy number and the change of the methylation site.It can be concluded that NAG18 is a nasopharyngeal carcinoma related gene located in the smallest common deletion region of 7q32 ter and is a highly conserved gene involved in the transcription of DNA activation