Abstract
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.
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Acknowledgements
We would like to thank Maria A.W.H. van Waarde for expert assistance on biochemical analysis. We thank Eefje Pelster, Alette H. Faber, and Reinier Bron for assistance on gene cloning. Lenja Bystrykh (Department of Cell Biology, Stem Cell Biology Section, University Medical Center Groningen, The Netherlands) is kindly acknowledged for help on the Unigene EST collection and Ron Dirks (section Biochemistry, Radbout University, Nijmegen, The Netherlands) for help on the Affimetrix gene array data. Russell S. Thomas (CIIT Centers for Health Research, NC, USA) is acknowledged for providing the array data sets. This work was supported by Innovatiegerichte Onderzoeksprogramma Genomics Grant IGE03018.
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Hageman, J., Kampinga, H.H. Computational analysis of the human HSPH/HSPA/DNAJ family and cloning of a human HSPH/HSPA/DNAJ expression library. Cell Stress and Chaperones 14, 1–21 (2009). https://doi.org/10.1007/s12192-008-0060-2
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DOI: https://doi.org/10.1007/s12192-008-0060-2