Encoding Gene Cloning and Expression Analysis of BYPASS1 in Brassica oleracea L.
LIU Yu-dong1,GAO Qi-guo1*,ZENG Jing2,ZHANG Lin-cheng1,ZHU Li-quan2,REN Xuesong1,WANG Xiao-jia1
(1College of Horticulture and Landscape Architecture, Southwest University, Key Laboratory of Horticulture Science
for Southern Mountainous Regions, Ministry of Education, Chongqing 400716, China; 2College of Agronomy
and Biotechnology, Southwest University, Chongqing 400716, China)
Abstract: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.