Cloning,Subcellular Localization and Expression Analysis of Heat Shock Protein Gene CpHSP70-1 from Chimonanthus praecox (L.)

Based on the EST analysis of Winter sweet transcriptome database constructed from Chimonan‐thus praecox flowers ,and using RACE technology ,the full‐length cDNA sequences of heat shock protein HSP70 was obtained by cloning ,named as CpHSP70‐1 (GenBank accession number :KR071130) .The full‐length of CpHSP70‐1 cDNA sequence was 2 520 bp ,with an open reading frame (ORF) of 1 962 bp , encoding a putative polypeptide of 653 amino acid residues and containing 3 signature motifs of the HSP70 family .CpHSP70‐1 protein was shown to have high homology with HSP70 proteins from other eu‐karyotes ,and phylogenetic analysis revealed that it belongs to the subclasses located in the nucleus/cyto‐plasm .Subcellular localization result supported the prediction that this protein is located in the nucleus . Real‐time fluorescence quantitative PCR was performed to analyze the expression characteristics of Cp H‐S P70‐1 gene .It was expressed ,at different levels ,in various tissues ,with the highest expression level found in mature leaves ;and sustained and stable expression was detected at different flower developmental stages .Cp H S P70‐1 gene expression increased under low or high temperature stresses ,and it responded more sensitively to high temperature .