作　　者： ; ; ; ; ; ; ; ; ;

机构地区： 中南大学湘雅医学院肿瘤研究所

出　　处： 《生物化学与生物物理进展》 2004年第5期464-469,共6页

摘　　要： 从UniGene库中选取编号为BG2 31197,来自人鼻咽组织的EST序列 .利用Blast检索GenBank的nr数据库和EST数据库 ,构建EST重叠群 .利用人类基因组草图搜索法从成人正常鼻咽组织中PCR扩增获得该基因全长cDNA ,命名为NAP1,GenBank登录号为AY190 32 6 .NAP1基因cDNA序列全长为 5 73bp ,编码由 85个氨基酸组成 ,相对分子质量为 970 0的多肽 .用α 3 2 P dCTP标记NAP1基因片段 ,与含 15种正常成人组织的多组织RNA印迹膜杂交 ,结果表明NAP1基因在淋巴结和气管中高表达 ,转录本大小约为 0 6kb ,在其他组织中不表达 .NAP1蛋白质与滤泡树突状细胞 (folliculardendriticcell,FDC)的一种分泌肽前体 (FDC SP) (AF4 35 0 80 )同源 ,与其他已知蛋白质无明显同源性 .NAP1基因定位在染色体 4q13,基因组跨越 9179bp ,含 5个外显子和 4个内含子 .采用差异RT PCR检测了 4 0例经病理诊断为低分化鳞状上皮癌的鼻咽活检组织及其对侧相应部位的正常鼻咽组织中该基因的表达差异 .在 4 0例鼻咽癌中 ,NAP1基因表达下调的有 17例 (4 2 5 % ) ,表达上调的有 6例 (15 % ) ,无明显表达差异的有 17例 (4 2 5 % ) .原位杂交和免疫组化结果显示该基因在正常鼻咽和鼻咽癌组织间质中的树突状细胞中表达 。 NPC-related ESTs were selected from UniGene bank. According to nr and EST database BLAST results, BG231197 was found to be a sequence that represents a new gene. The EST contig does not have complete ORF. NAP1 gene cDNA was got from normal nasopharynx tissue cDNA by PCR through searching Human Genome Draft. The Northern blot analysis revealed one transcript (0.6 kb) in lymph node and trachea. GenBank Accession number of the gene is AY190326. The length of NAP1 gene cDNA is 573 lip, ORF is from 113 bp to 370 lip. NAP1 encodes a 9 700 u protein composed of 85 amino acids. The protein resembles to FDC-SP secreted by FDC (AF435080). Bioinformatics analysis suggested that the protein is a secreted protein. NAP1 gene is located at chromosome 4q13. Genome spans 9 179 lip, containing 5 exons and 4 introns. The difference of NAP1 expression between nasopharyngeal. carcinoma ( NPC) and nasopharynx (NP) tissues of 40 cases was tested by differential RT-PCR. Low expression occurred in 17 NPC cases. High expression occurred in 6 NPC cases. Expression difference did not occur in 17 NPC cases. In situ hybridization ( ISH) and immunohistochemistry (III) suggested that it express in dentritic cells of NP and NPC tissues. It is meaningful to do further studies on the reason of low expression in NPC tissue and the function of NAP1 gene.

关 键 词： 鼻咽癌 基因 基因克隆 鉴定

领　　域： [生物学]