USE OF A FAST PROTEIN ELECTROPHORETIC PURIFICATION PROCEDURE FOR N-TERMINAL SEQUENCE-ANALYSIS TO IDENTIFY S-LOCUS RELATED PROTEINS IN STIGMAS OF BRASSICA-OLERACEA

被引:23
作者
GAUDE, T
DENOROY, L
DUMAS, C
机构
[1] UNIV LYON 1, INRA, RCAP, F-69622 VILLEURBANNE, FRANCE
[2] CNRS, SERV CENT ANAL, F-69390 VERNAISON, FRANCE
关键词
D O I
10.1002/elps.1150120909
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In the cruciferous plant Brassica oleracea L. (cabbage), the S-locus specific glycoproteins (SLSGs) isolated only in stigmas are considered to play an important role in the normal prevention of self-fertilization. Recent molecular data have shown that the gene encoding these glycoproteins (the SLG gene) belonged to a multigenic family consisting of about 10 homologous copies among which another member is expressed, the S-locus related gene (SLR1gene). Our aim was to determine whether the SLR1-gene proteins were expressed in the stigmatic tissues. We first identified the putative SLSGs or SLR1-proteins by Con A-peroxidase detection of glycoproteins separated after isoelectric focusing in polyacrylamide gels. We describe a fast purification procedure for the glycoproteins of interest, based on analytical isoelectric focusing, electrophoresis, and electroblotting of proteins onto polyvinylidene difluoride membranes. Blotted proteins were sequenced for N-terminal amino acid determination. By comparison of the N-terminal sequences of the purified proteins with the peptide sequence predicted from the SLR1-cDNA, we demonstrate the expression of SLR1-like proteins in stigmas of B. oleracea.
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收藏
页码:646 / 653
页数:8
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