摘要
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 。
In order to clone a novel putative NPC associated gene on the smallest common deletion region of 7q32-ter. BAC clone was screened by PCR using STS D7S509 probe. The up-regulated expression of 3' end expressed sequence tags (ESTs) localized within this smallest common deletion region were screened in NPC cell line HNE1 and NPC biopsies using EST-mediated positional candidate clone and bioinformatics. The full-length cDNA of candidated EST was cloned through cDNA clone sequencing and bioinformatics. Southern blot and methylation analysis were used to study the machanism of up-regulated expression of NAG18 in NPC. The results showed that the full-length cDNA of NAG18 is 802bp, its encoding protein is 227 amino acids. It is highly homologous to human and mouse TAXREB107 and RPL6. Loss of gene copies and aberrant methylation are not the machanism of its up-regulated expression. It can be concluded that the gene NAG18 located in this region may be a putative NPC associated gene. It is a highly conserved gene. It may participate in Tax-mediated tran-activation of transcription.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2001年第5期711-716,共6页
Progress In Biochemistry and Biophysics
基金
国家 8 63项目 (10 2 10 0 1 0 5 )
973重大项目 ("疾病基因组学"理论和技术体系的建立鼻咽癌子项目 )资助~~
关键词
7q32-ter最小共同缺失区
鼻咽癌
相关基因
染色体
nasopharyngeal carcinoma
the smallest common deletion region of 7q32-ter
NPC associated gene
