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利用减法杂交和RACEs从水稻花中克隆了1个编码含丰富脯氨酸和苏氨酸结构域多肽的cDNA,其相应的基因命名为RA68.RA68含3个外显子和2个内含子,编码由219个氨基酸残基组成的蛋白质.数据分析结果显示,该蛋白由1个22个氨基酸残基组成的信号肽,1个亲水性的N-端结构域和1个疏水性的C-端结构域组成.N-端结构域是一段嵌合PTPTSYGmotif的富含脯氨酸和苏氨酸的序列.Southernblot和序列分析结果表明,RA68在水稻基因组以单拷贝存在,定位于第2号染色体.Northern杂交结果表明,RA68在幼芽和花中表达量较高,在根和叶中不表达.花和幼芽进行原位杂交分析结果表明,在花中,RA68在花粉母细胞、二分体细胞、单核花粉粒以及大孢子囊中表达.基于该基因的表达特性及其编码蛋白质的结构特征,探讨了其在花发育中的可能功能.
A cDNA encoding a polypeptide rich in proline and threonine was cloned from rice flower using subtractive hybridization and RACEs, and the corresponding gene was named RA68. RA68 contains 3 exons and 2 introns , Encoding a protein consisting of 219 amino acid residues.Data analysis showed that the protein consists of a signal peptide consisting of 22 amino acid residues, a hydrophilic N-terminal domain and a hydrophobic C - terminal domain. The N-terminal domain is a chimeric PTPTSYGmotif which is rich in proline and threonine. Southern blot and sequence analysis showed that RA68 is present in a single copy of the rice genome and is located on the 2nd The results of Northern hybridization showed that the expression of RA68 was higher in shoots and flowers and not in roots and leaves.The in situ hybridization analysis of flowers and shoots showed that RA68 was expressed in pollen mother cells, Segregating cells, mononuclear pollen grains and megaspore vesicles.According to the expression characteristics of the gene and the structural characteristics of its encoded protein, the possible functions of the gene in flower development were discussed.