Three full-length cDNA sequences of PGK genes(BnPGK1,BnPGK2 and BnPGK3) were obtained through electronic cloning method based on bioinformatics analysis of Arabidopsis PGK gene family for the study of Phosphoglycerate kinase(PGK) gene expression characteristics from Brassica napus.Specific primers were designed and cDNA of male sterile line 09A and maintainer line 09B were as templates to clone BnPGK full-length sequences.Real-time fluorescence quantitative specific primers were designed according to the obtained cDNA sequences.The Real-time quantitative analysis was conducted by using Quantitative PCR method.Results showed that the BnPGK genes expressed variously in different tissues,such as root,stem,leaf and bud of 09A and 09B,belong to constitutively expressed.Except BnPGK3 in the stem,the expression of BnPGK other genes of 09A were higher than that of 09B in root,stem and leaf,but the expression of BnPGK genes of 09B were higher than that of 09A and the expression of BnPGK1 and BnPGK3 of 09B was 2 times than that of 09A in bud.