脑胶质瘤患者全长新基因的获得及初步研究

目的 探讨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初步研究。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507E08克隆子进行了Northern印迹、原位杂交、生物信息学分析和染色体定位。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Northern印迹证实507E08基因在人正常脑组织中低表达，而在人脑胶质瘤中高表达。原位杂交得到相同的结果。BLASTn和BLASTx分析显示，507E08基因为全长新基因，长度为2002bp，共编码203个氨基酸。本基因命名为人核糖体蛋白L14．22。该基因定位于14号染色体上D14S1066和D14S265Marker之间。结论 用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。507E08基因可能是与人脑胶质瘤形成有关的一条全长新基因。

Isolation and studv of one novel full-length gene related to human glioma

Objectives To obtain differentially expressed genes related to human glioma using cDNA microarray and make a preliminary study 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 named 507E08 clone was subsequently analyzed by northern blotting, in situ hybridization, bioinformatics and radiation hybridization. Results Fifteen differentially expressed novel genes related to human glioma were obtained through four times of hybridization and scanning. Northern blotting and in situ hybridization confirmed that 507E08 clone was lowly expressed in normal human brain tissue and over expressed in human glioma tissue. BLASTn and BLASTx analysis showed that the clone 507E08 was a novel full length gene with the length of 2002 bp. This gene, called human ribosomal protein 14.22 gene, codes 203 amino acids and is located on chromosome 14 between D14S1066 Marker and D14S265 Marker. Conclusion cDNA microarray technology can be successfully applied to identify differentially expressed genes with small amount of specimen, high quality, high speed, and high sensitivity. The novel full length human ribosomal protein 14.22 gene may correlate with formation of human glioma.