Encoding Gene Cloning and Expression Analysis of BYPASS1 in Brassica oleracea L.
An up-regulated protein in Brassica oleracea L.stigma induced by self-pollination was identified as BYPASS1 protein by 2-DE electrophoresis and MALDI-TOF-TOF/MS.The genomic DNA and cDNA coding sequences of BYPASS1 (BPS1) were amplified by PCR.The sequence analysis showed that BPS1 had no intron.Its ORF was 1 059 bp,encoded a polypeptide with 352 amino acids with a predicted molecular mass of 38.7 kD.Phylogenetic tree analysis using MEGA 5.1 showed that Brassica oleracea L.BPS1 was more close to Arabidopsis thaliana BPS1 rather than Nicotiana benthamiana BYPASS1,amino acid homologous similarity is 85％.RT-PCR analysis showed that 1-2 days before flowering,BPS1 expressed in petal,sepal,pollen,stigma and leaf,and the express level in the stigma was higher than the other organs.Real-time fluorescence quantitative PCR analysis showed that the relative expression level of BPS1 in stigma was continually increased after self-pollination,and after cross-pollination its expression level was increased firstly,than decreased and finally increased again.
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