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cDNA microarray in isolation of novel differentially expressed genes related to human glioma and clone of a novel full-length gene
Authors:Qi Zhen-yu  Hui Guo-zhen  Li Yao  Zhou Zong-xiang  Gu Shao-hua  Ying Kang  Xie Yi
Institution:1. Department of Neurosurgery, First Affiliated Hospital of Soochow University, Suzhou 215006, China
2. State Key Laboratory of Genetic Engineering, Institute of Genetics, Fudan University, Shanghai 200433, China
Abstract:BACKGROUND: This investigation was undertaken 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, the results were scanned using a computer system. The gene named 681F05 clone was an expressed gene to human glioma through four-time hybridization and scanning. Subsequently northern blot analysis was performed by northern blot, 5'RACE and bioinformatics. RESULTS: Fifteen differentially expressed genes to human glioma were obtained through four-time hybridization and scanning. Northern blot analysis confirmed that 681F05 clone was low-expressed in human brain tissues and over-expressed in human glioma tissues. The analysis of BLASTn and BLASTx showed that 681F05 clone is two cDNA clones encoding two novel proteins that are highly identified to the cyclophilin isoform 10 of C. Elgans, respectively. Sequence analysis revealed the two cDNA clones are two different splicing variants of a novel cycophilin-like gene (PPIL3a and PPIL3b). CONCLUSIONS: cDNA microarray technology can be successfully used to identify differentially expressed genes. The novel full-length gene of human PPIL3 may be correlated with the formation of human glioma.
Keywords:cDNA microarray  differential expression  glioma  novel full-length gene
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