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目的运用基因芯片技术获取正常成人脑组织与人脑胶质瘤中差异表达的基因,并对其中一条基因进行了初步的研究。方法抽提正常成人脑组织与人脑胶质瘤组织中的mRNA来制备探针,经杂交、洗涤后,通过计算机观察两者表达谱的差异情况,对681F05克隆子进行了Northernblot,5’RACE和生物信息学分析。结果通过4次基因芯片筛选,获得15条与胶质瘤相关的新基因,经Northernblot证实681F05基因在人正常脑组织中低表达,而在人脑胶质瘤中高表达。BLASTn和BLASTx分析显示,它们编码蛋白与线虫Cyp10蛋白同源性分别为52%和72%。cDNA序列分析发现这两上克隆是同一个基因[命名为cyclophilinlikegene(PPIL3)]的两个不同的剪切体(PPIL3a和PPIL3b)。结论基因芯片筛选正常脑组织与人脑胶质瘤差异表达的基因具有样品用量少,高质量,高速度,高敏感等特性。681F05基因可能是与人脑胶质瘤形成有关的一条全长新基因。
OBJECTIVE: To obtain genes differentially expressed in normal adult human brain and human glioma using gene chip technology and to study a gene. Methods The mRNA was extracted from normal adult human brain and human glioma tissue. After hybridization and washing, the expression of 681F05 was observed by computer. Northernblot, 5’RACE And bioinformatics analysis. Results Fifteen novel genes related to glioma were obtained by four microarray analysis. Northern blot showed that the 681F05 gene was low expressed in normal human brain tissue and highly expressed in human glioma. BLASTn and BLASTx analyzes showed that the homology of their coding proteins to the nematode Cyp10 protein was 52% and 72%, respectively. cDNA sequence analysis revealed that the two clones were two different splicers (PPIL3a and PPIL3b) of the same gene [named cyclophilinlikegene (PPIL3)]. Conclusion Gene microarray screening of normal brain tissue and human glioma differentially expressed genes with less sample amount, high quality, high speed, high sensitivity and other characteristics. The 681F05 gene may be a new full-length gene associated with human glioma formation.