摘要
对蛋白质组学的研究有许多不同的切入方法 .从研究的生物学意义和可行性考虑 ,提出从蛋白结构域入手进行蛋白质组学研究 .SH2 (Srchomology 2 )结构域是细胞信号转导中重要的元件之一 ,人SH2结构域共有约 12 0种 ,对其进行研究将深刻揭示细胞信号转导的规律 .为了得到人所有的SH2结构域序列及克隆 ,首先在公共数据库里检索出了人所有的SH2结构域序列 ,利用国际上现有的共享资源IMAGE(IntegratedMolecularAnalysisofGenomesandTheirExpression)克隆为PCR模板 ,解决了从cDNA文库中难以克隆低丰度结构域的问题 .利用有方向性的TOPO克隆技术提高克隆效率 ,从而快速高效地构建了包括 6 0个SH2结构域的克隆库 .克隆库可以方便地转换到GATEWAY系统具有各种用途的载体上 。
There are many approaches for proteomics research. It was proposed to study domains in proteomics because domains were biologically important and members of each domain were limited. SH2 (Src homology 2) domain was an important element in signal transduction and there were about 120 SH2 domains in human cells. Public databases were searched to obtain all human SH2 domain sequence information. Royalty-free IMAGE clones were used as template instead of cDNA library for high efficiency directional TOPO cloning into multipotential GATEWAY entry vector. All clones were verified by sequencing. A SH2 domain library with sixty clones was constructed rapidly for SH2 domain proteomics research.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2003年第4期537-541,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家高技术研究发展计划 (863 ) (No .2 0 0 1AA2 3 3 0 5 1)
国际科技合作重点项目计划 (No .2 0 0 2AA2 2 90 3 1)
重大基础研究前期研究专项项目计划(No .2 0 0 2CCA0 410 0 )资助~~
