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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 ,这为脑胶质瘤的基因治疗提供了一条新思路
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 of normal human brain tissue and human glioma tissue was extracted to prepare probes. After hybridization and washing, the 436F11 clones were analyzed by northern blot and in situ hybridization And bioinformatics analysis. Results Fifteen new genes related to glioma were obtained by four microarray analysis. Northernblot showed that 436F11 gene was highly expressed in human normal brain tissue and low in human glioma. In situ hybridization yielded the same result. BLASTn and BLASTx analysis showed that the 436F11 gene was a new full-length gene encoding a total of 78 amino acids. The theoretical molecular weight was 86 48Da and the isoelectric point was 469. The 436F11 gene was homologous to the 9% homolog of PKIβ and named as human PKIβ. 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. Human PKIβ may be a new full-length gene associated with human glioma formation, which provides a new idea for gene therapy of glioma