CAJ | 학술논문

目的应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1条基因进行初步研究.方法抽提正常成人脑组织与人腩胶质瘤组织中的mRNA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,对681 F05克隆子进行Northern blot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Northern blot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Cyp-10蛋白同源性分别为52%和72%.cDNA序列分析发现此两个克隆是同一个基因[命名为cyclophilin-like gene(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.
목적 응용기인심편기술획취정상성인뇌조직여인뇌효질류중차이표체적기인,병대기중1조기인진행초보연구.방법 추제정상성인뇌조직여인남효질류조직중적mRNA제비탐침,경잡교、세조후,통과계산궤관찰량자표체보적차이,대681 F05극륭자진행Northern blot화생물신식학분석.결과 통과4차기인심편사선,획득15조여효질류상관적신기인,경Northern blot증실681F05기인재인정상뇌조직중저표체,이재인뇌효질류중고표체.BLASTn화BLASTx분석현시,타문편마단백여선충Cyp-10단백동원성분별위52%화72%.cDNA서렬분석발현차량개극륭시동일개기인[명명위cyclophilin-like gene(PPIL3)]적량개불동적전절체(PPIL3a화PPIL3b).결론 기인심편사선정상뇌조직여인뇌효질류차이표체적기인구유양품용량소、고질량、고속도、고민감등특성.681F05기인가능시여인뇌효질류형성유관적일조전장신기인.
Objective To obtain differentially expressed genes related to human glioma using cDNA microarray and study 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 named 681F05 clone was subsequently analyzed by northern blotting and bioinformatic analysis. Results We obtained 15 differentially expressed genes to human glioma through 4 times hybridizations and scanning. Northern blotting analysis confirmed 681F05 clone was down-regulated in human brain tissue and up-regulated in human glioma tissues. The analysis of BLASTn and BLASTx showed that clone 681F05 isolated was two cDNA clones encoding two novel proteins which showed 52% and 72% identity to the cyclophilin isoform 10 of C. Elgans, respectively. Sequence analysis revealed these two cDNA clones were two different splicing variants of a novel cycophilin-like gene ( PPIL3a and PPIL3b). Conclusion cDNA microarray technology can be successfully applied to identify differentially expressed genes. The novel fulllength gene of human PPIL3 may be correlated with tumorigenesis of human glioma.