重金属暴露铜锈环棱螺全组织均一化cDNA文库构建及表达序列标签特征分析
Construction of Normalized cDNA Library and Analysis of Expressed Sequenced Tags Properties of Bellamya Aeruginosa
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摘要: 构建cDNA文库是寻找生物新功能基因的有效手段。采用SMART技术构建了重金属铜和镉暴露铜锈环棱螺全组织均一化cDNA文库,文库库容为1.78×106克隆,重组率大于99%。从初始文库中随机挑取6000个克隆进行测序,获得5473个高质量表达序列标签(ESTs)序列。经聚类分析得到3961个Unigene(平均长度为815bp),其中包含897条重叠群(Contigs),3604条单拷贝EST序列(Singlets),冗余度为27.63%。对ESTs序列进行相似性功能分类(COG)、标准基因词汇体系(GO)功能分类和基因与基因组(KEEG)代谢途径的生物信息学分析。结果表明此文库含有大量的生长发育、细胞信号传导机制和生物防御机制等相关基因,为进一步筛选铜锈环棱螺功能基因奠定了生物信息学基础,也为发展铜锈环棱螺毒理效应研究提供了丰富的信息资源。Abstract: Construction of normalized cDNA library is an available tool for identifying novel genes of organisms.In the present study,a cDNA library of Bellamya Aeruginosa exposed to heavy metals through SMART technique was constructed.The tier of cDNA library was 1.78×106 plaques,with a recombinant rate of more than 99%.6 000 clones selected randomly and 5 473 expressed sequenced tags(ESTs) were obtained.After processing,a total of 3 961 unigenes comprising 897 contigs and 3 064 singlets were obtained,and the redundancy was 27.63%.Analysis of EST using COG,GO and KEGG showed that a large group of genes related to growing development,signal transduction mechanisms of cells,defense mechanisms were obtained in the present cDNA library.The normalized cDNA library developed in the present study provides a valuable resource for further research on functional genes and ecotoxicology to Bellamya Aeruginosa.
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