in this study, we analyzed the differential proteome of winter rapeseed (Brassica rapa), the Two-dimensional gel electrophoresis (2D-DIGE) technology combined with the Liquid Chromatography-Mass Spectrometry (LC-MS) technology to detect the changes of differential proteome in DR-5 (drought-resistant Winter Rapeseed line) under drought stress, cloned the heat shock cognate protein 70 gene (HSC70-1) and analyzed the expression of this gene under drought stress. In order to provide a theoretical base for drought resistance breeding of winter rapeseed, the effect of moderate drought stress on winter rapeseed was studied. We obtained the coding region sequence of HSC70-1 in winter rapeseed by homologous cloning, and analysed the physicochemical properties, secondary structure, tertiary structure, signal peptide, transmembrane structure, conserved region, active pocket and phylogenetic conservatism of this gene encoding protein HSC70 by bioinformatics analysis methods. The Real-time fluorescence quantitative and semi-quantitative analysis found the changes of HSC70-1 gene expression levels under drought stress. There were 23 differentially expressed proteins successfully identified by Mass spectrometry, which were involved in stimulation response (5), glucose/energy metabolism (6), lipid metabolism (2), signal transduction (1), amino acid/protein metabolism (2), nucleic acid metabolism (1), cytoskeleton (1), photosynthesis (2) and chaperone protein (3). We obtained the 1 911 bp complete open reading frame of HSC70-1 gene via cloning, which encoded 637 amino acid sequences. HSC70 is a stable hydrophobic protein, and it contains the unique nucleic acid and substrate binding functional area of HSP protein family, it have four active pockets. The amino acid sequence was highly similar to HSC70 in Brassica rape, and the similarity was 98%, which is highly conserved. The expression of HSC70-1 in winter rapeseed leaves was up-regulated under drought stress, which improved the tolerance of drought stress in winter rapeseed.