三叶木通花粉前纤维蛋白基因的克隆与表达

以三叶木通花蕾为材料,采用RT-PCR、3-′RACE方法克隆了三叶木通花粉前纤维蛋白基因,命名为Atf-Pro(GenBank登录号GQ478584)。结果表明:AtfPro的cDNA全长735 bp、阅读框393 bp、编码131个氨基酸,有1个342 bp的3′端非翻译区。预测分子量约为14.081 kD,等电点4.74。氨基酸和核苷酸序列的同源性分析发现,AtfPro基因属于植物花粉profilin基因家族的新成员。RT-PCR定性分析表明,AtfPro基因在三叶木通花蕾、花药、雌花花瓣和柱头组织中均有表达,但在幼叶、茎尖、根尖组织中低水平表达或不表达,生殖器官中的表达时期从花序分化发育开始到开花散粉结束。

Cloning and Expression Analysis of a Pollen Profilin Gene from Akebia trifoliata

With Akebia trifoliata bud as material,RT-PCR and 3′-RACE was used to clone the pollen profilin gene.A gene coding for this pollen profilin,designated as AtfPro(GenBank accession number:GQ478584) was isolated from Akebia trifoliata bud.The full-length AtfPro cDNA is 735 bp,an ORF of 393 bp,and a 342 bp 3′ UTR.This cDNA sequence encoded a polypeptide of 131 amino acid residues with a predicted molecular mass of 14.081 kD and a basic isoelectric point of 4.74.The AtfPro gene belongs to pollen profilin family through the nucleic acid and amino acid sequence homology analysis.Qualitative RT-PCR analysis showed that AtfPro gene were expressed in reproductive organs such as trifoliate bud,male flower anthers,female petals and stigma,but low level of expression or no expression in vegetative organs,such as young leaves,shoot and root tip.The expression reproductive organs include the process from the inflorescence differentiation to flower shedding.