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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 ,利用生物信息学技术分析克隆基因。方法:采用 SMART方法构建 c DNA文库 ,杂交筛选并克隆基因 ,利用生物信息学手段对克隆基因进行结构和功能的分析。6 μm ol/ L Na As O2 处理人正常肝 L- 0 2细胞 2 h,抽提 RNA做基因芯片杂交。结果:成功克隆一条新基因 ,生物信息学分析发现在 7到 4 5 6碱基范围内有一个完整的 ORF,在编码起始区有典型的 Kozak序列 ,编码 14 9个氨基酸的蛋白质。核酸同源性比对发现与人类 1p36 .2 - 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:克隆的人类砷相关基因 ,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 c DNA library was constructed by SMART method, and the genes were screened and cloned by hybridization. The structure and function of the cloned genes were analyzed by bioinformatics methods. Human normal liver L-02 cells were treated with 6 μmol / L Na As O2 for 2 h, and RNA was extracted for gene chip hybridization. Results: A new gene was successfully cloned. Bioinformatics analysis showed that there was a complete ORF in the range of 7 to 456 nucleotides, a typical Kozak sequence in the coding region and a protein of 149 amino acids. Nucleotide homology comparison revealed 98% homology with human 1p36.2-36.3 chromosomal DNA sequence and 85% homology with Alu subfamily SB sequence. Dye arsenic cell gene chip hybridization found that cloned gene expression increased. Conclusion: The cloned human arsenic related genes have high homology with the Alu subfamily SB sequence.