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利用cDNA-AFLP技术分析大白菜核雄性不育两用系‘AB02’可育株(msms)和不育株(Msms)花蕾的基因表达谱,在可育株混合花蕾cDNA中扩增出1条特异条带TDF-25,通过RACE和RT-PCR技术克隆了该基因的全长cDNA序列。序列分析表明,该基因编码脂质转移蛋白,命名为BrLTP1。BrLTP1全长cDNA序列为750bp,推测编码1个包含183个氨基酸残基的前体蛋白。BrLTP1蛋白含有典型的脂质转移蛋白N端信号肽,保守的AAI结构域和半胱氨酸位点。预测BrLTP1蛋白含有多种修饰性位点,包括1个PKC磷酸化位点,2个N-糖基化位点和10个N-端豆蔻酰基化位点。基因表达模式表明,BrLTP1在两用系不育株花蕾中受到强烈抑制,在可育株的大花蕾、成熟花药以及花瓣中高水平表达。
CDNA-AFLP was used to analyze the gene expression profiles of flower buds of MSMS male sterile lines (ABMS ’msms) and sterile plant (Msms) in Chinese cabbage. One The full-length cDNA sequence of TDF-25 was cloned by RACE and RT-PCR. Sequence analysis showed that the gene encoding lipid transfer protein, named BrLTP1. The full-length cDNA sequence of BrLTP1 is 750bp, presumably encodes a precursor protein containing 183 amino acid residues. The BrLTP1 protein contains the typical lipid transfer protein N-terminal signal peptide, the conserved AAI domain and the cysteine site. The BrLTP1 protein is predicted to contain a variety of sites for modification, including one PKC phosphorylation site, two N-glycosylation sites and ten N-terminal myristoylation sites. The pattern of gene expression indicated that BrLTP1 was strongly inhibited in the flower buds of both lines of sterile plants and expressed at high levels in large flower buds, mature anthers and petals of fertile plants.