采用基因芯片技术获取人脑胶质瘤差异表达的基因及一条全长新基因的研究

Isolation of differentially expressed genes related to human glioma using cDNA microarray and the study of one novel full-length gene

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 ,通过计算机观察二者表达谱的差异情况 ,对 436F11克隆子进行了northernblot,原位杂交和生物信息学分析。结果 通过四次基因芯片筛选 ,获得 15条与胶质瘤相关的新基因 ,经northernblot证实 436F11基因在人正常脑组织中高表达 ,而在人脑胶质瘤中低表达。原位杂交得到相同的结果。BLASTn和BLASTx分析显示 ,436F11基因为全长新基因 ,共编码 78个氨基酸 ,其理论分子量为 86 48Da ,等电点为 4 6 9,与鼠PKIβ 6 9%同源 ,命名为人PKIβ。 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。人PKIβ可能是与人脑胶质瘤形成有关的一条全长新基因 。

Objective To obtain differentially expressed genes related to human glioma using cDNA microarray and the characterization of one novel full length gene.Methods Total RNA was extracted from human glioma tissues and normal brain tissues, and mRNA was used to make probes.After hybridization and washing procedure,the results of hybridization were scanned using computer system.One gene naned 436F11 clone was subsequently analyzed by northern blot,in situ hybridization and bioinformatics.Results We obtain 15 differentially expressed genes to human glioma thruogh four times hybridizations and scanning.Northern blot analysis confirmed 436F11 clone was over expression in human brian tissue and low expression in human glioma tissues.This result is just the same as the result of in situ hybridization.The analysis of BLASTn and BLASTx showed that clone 436F11 was a novel full length bene.This gene codes 78 amino acid of protein,whose theoretical molecular weight is 8648 Da and PI is 4 69,while it is 60% identity to mouse PKIβ amino acid, so it is called human PKIβ gene.Conclusions We find that cDNA microarray technology can be successfully applied to identify differentially expressed genes.The novel full length gene of human PKIβ may be correlate with forming of human glioma,and it could provide a new way to gene therapy of glioma.