Cloning And Expression Of HSF1 CDNA From Hainan Eld’s Deer

The investigation of heat shock transcription factor 1 has benefit for interpretation the molecular mechanism of the heat shock response,with significant theoretical value. Full length of HSF1 cDNA of Hainan Eld’s Deer-hdHSF1-was amplified by RT-PCR and RACE, and cloned into pMD20-T vector. The recombinant plasmid was confirmed by endonuclease digestion、sequencing. The PCR product was then subcloned into pET28a vector and transformed into E.coli host cells. Protein expression was induced by IPTG and analyzed by SDS-PAGE and Western blot. The results showed that the hdHSF1 cDNA was 2036 bp in length and contained a 1578 bp ORF encoding 525 amino acids and the encoded protein was a hydrophilicity protein which Ip is 4.93. The homology of HSF1 gene of Hainan Eld’s Deer compared to that of cattle、mouse and human are 96.20％、88.39％、81.53%, and the homology comparison of HSF1 amino acid sequence on Hainan Eld’s Deer with cattle、human and mouse was 97.52%、89.25％、83.84% respectively. Phylogenetic tree based on the HSF1 protein of different animals was matched to the traditional taxonomic classification of the animal. A 62kD fusion protein with a His tag was induced by IPTG and confirmed by Western blot using anti-His·Tag monoclonal antibody, getting a special antibody binding band.These results indicated that prokaryotic expression vector of hdHSF1 was constructed and expressed successfully.