甘蓝SRK底物蛋白基因的克隆与序列分析

采用PCR、3’RACE以及其他分子生物学方法,以甘蓝基因组DNA和柱头cDNA为模板对SSP基因进行扩增克隆,得到长度分别为778 bp和825 bp的基因片段。序列分析表明,甘蓝SSP基因不包含内含子,SSP核苷酸序列与拟南芥肽酶M28家族蛋白有同源性,序列相似性达到67.4%,两者的氨基酸序列相似性达到62.7%,而且都含有一个TFR_dimer结构域,推测SSP蛋白极有可能与在自交不亲和过程中受ARC1作用而泛素化了的某一未知蛋白的降解相关。

Cloning and Sequences Analysis of SRK Substrate Protein Gene from Brassica oleracea

The DNA and cDNA fragments of SRK Substrate Protein Gene were initially amplified from genomic DNA and stigma cDNA in Brassica oleracea by using PCR,3'RACE and other molecular method.Their lengths were 778bp and 825 bp.Sequence analysis indicated that the SSP gene may has no intron.We also found that SSP from Brassica oleracea shared high similarity with the gene sequence of Arabidopsis thaliana peptidase M28 family protein up to 67.4% identity.Homologous analysis in amino acid sequence showed that between SSP and Arabidopsis thaliana peptidase M28 family protein were 62.7% in identity.SSP contained a TFR_dimer domain(Transferrin receptor-like dimerisation domain)which also exists in Arabidopsis thaliana peptidase M28 family proteins.We presumed that SSP is associated with the degradation of an unknown protein initiated by ARC1 in SSI signal transduction process.