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目的探讨砷化物与基因的相互作用,克隆砷相关基因,利用生物信息学分析克隆基因。方法利用SMART方法构建cDNA文库,杂交筛选并克隆基因,生物信息学对克隆基因进行结构和功能的分析。6μmol/LNaAsO2处理人正常肝L鄄02细胞2h,抽提RNA做基因芯片杂交。结果成功克隆一条新cDNA基因,生物信息学分析发现7到456有一个完整的ORF,在编码起始区有典型的Kozak序列,编码149个氨基酸的蛋白质。核酸同源性比对发现与人类1p36.2鄄36.3染色体DNA序列同源性达98%,编码蛋白质同Alu亚家族SB序列同源性达85%。染砷细胞基因芯片杂交发现克隆基因表达增高。结论克隆了人类砷相关基因,该基因编码蛋白质与Alu亚家族SB序列同源性高。
Objective To investigate the interaction between arsenic and gene, clone arsenic related gene and analyze the cloned gene by bioinformatics. Methods The cDNA library was constructed by SMART method, and the genes were selected by hybridization and cloned. The structure and function of the cloned genes were analyzed by bioinformatics. Human normal L-02 cells were treated with 6μmol / L NaAsO2 for 2h, RNA was extracted for gene chip hybridization. Results A new cDNA gene was successfully cloned. Bioinformatics analysis revealed that there was a complete ORF from 7 to 456 with a typical Kozak sequence encoding a 149 amino acid protein. Nucleotide homology comparison revealed 98% homology with the human 1p36.2-36.3 chromosomal DNA sequence, and the homology of the encoded protein to the Alu subfamily SB sequence was 85%. Dye arsenic cell gene chip hybridization found that cloned gene expression increased. Conclusion Human arsenic related gene was cloned, which encodes a protein with high homology to Alu subfamily SB sequence.