Hageman Jurre, Kampinga Harm H
Section of Radiation and Stress Cell Biology, Department of Cell Biology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
Cell Stress Chaperones. 2009 Jan;14(1):1-21. doi: 10.1007/s12192-008-0060-2. Epub 2008 Aug 7.
In this manuscript, we describe the generation of a gene library for the expression of HSP110/HSPH, HSP70/HSPA and HSP40/DNAJ members. First, the heat shock protein (HSP) genes were collected from the gene databases and the gene families were analyzed for expression patterns, heat inducibility, subcellular localization, and protein homology using several bioinformatics approaches. These results can be used as a working draft model until data are confirmed by experimental approaches. In addition, we describe the generation of a HSPA/DNAJ overexpression library and tested the effect of different fusion tags on HSPA and DNAJ members using different techniques for measuring chaperone activity. These results show that we have cloned a high-quality heat shock protein expression library containing most members from the HSPH, HSPA, DNAJA and DNAJB families which will be useful for the chaperone community to unravel the function of the highly diverse family of human molecular chaperones.
在本手稿中，我们描述了用于表达HSP110/HSPH、HSP70/HSPA和HSP40/DNAJ成员的基因文库的构建。首先，从基因数据库中收集热休克蛋白（HSP）基因，并使用多种生物信息学方法分析基因家族的表达模式、热诱导性、亚细胞定位和蛋白质同源性。在通过实验方法确认数据之前，这些结果可作为工作草案模型。此外，我们描述了HSPA/DNAJ过表达文库的构建，并使用不同的技术测量伴侣活性，测试了不同融合标签对HSPA和DNAJ成员的影响。这些结果表明，我们已经克隆了一个高质量的热休克蛋白表达文库，其中包含来自HSPH、HSPA、DNAJA和DNAJB家族的大多数成员，这将有助于伴侣蛋白研究领域解析人类分子伴侣这一高度多样化家族的功能。
