藏族外周血淋巴细胞表达序列标签分析

Analysis of expressed sequence tags of cDNA library from peripheral blood lymphocyte of Tibetan

目的对藏族外周血淋巴细胞cDNA文库进行随机大规模表达序列标签(ESTs)测序并进行生物信息学分析,通过序列比对找到与低氧适应相关基因有关的单核苷酸多态性(SNPs)位点。方法将部分文库经大肠杆菌BM 25.8质粒化,用5′端测序引物进行测序,利用BLAST程序对获得的ESTs从核苷酸和氨基酸水平进行同源性搜索和比对。结果经序列比对发现突变位点较多发生在UTR区域,同时一些基因的编码区域也发生蛋白序列的变化。对这些基因注释后发现,有些突变发生在与能量代谢、线粒体相关的基因及一氧化氮合酶上,如GTPase、NDUFA6、ATP5L、NOSIP。结论 ESTs技术是一个快速、有效寻找新基因的方法 ,是系统研究不同人群基因表达模式的有力工具。

Objective Sequencing large-scale expressed sequence tags randomly in the cDNA library from peripheral blood lymphocyte of tibetan and taking an analysis of bioinformatics,to find related SNPs sites of adaptation to hypoxia genes according to sequence contrast.Methods The partial library from E.coli BM 25.8 plasmids was sequenced using 5′ sequencing primer,and was made homology analysis and sequence contrast with the expressed sequence tags by BLAST procedure.Results It was found by sequence contrast that some mutation sites occurred mainly on UTR region,and a lot of protein sequences changed according to changes of genes coding region.After making clear these genes,we found that some mutations occurred on these genes were relative to energy metabolize,mitochondrion and nitrous oxidease,such as GTPase,NDUFA6,ATP5L,NOSIP.Conclusion Expressed sequence tags technique was a quick and efficient approach to find new genes and powerful tool to study gene expression model of different population.