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利用RT-PCR方法从矮牵牛(Petunia hybrida)中克隆了MADS盒基因pMADS4,进行了全序列测定。测序结果显示,所得到的基因与发表的序列同源率为100%。该基因长度为910bp,含有一个开放阅读框,编码253个氨基酸残基组成的蛋白,具有典型的植物MADS盒基因的结构,该蛋白序列与玉米(Zeamays)ZAG3、麻竹(Dendrocalamus latiflorus)DlMADS18和拟南芥(Arabidopsis thaliana)AGL6的同源性分别为64%、64%和61%,说明pMADS4是AGL6在矮牵牛中的同源基因,推测它们具有相似的功能,都促进开花并且调节花器官形成。进化树分析显示,AP1/AGL9亚家族分为SEP、AGL6和AP1三个分枝,且AGL6分枝又进一步分为三组,分别与裸子植物、单子叶植物和双子叶植物类群相对应,预测通过分离植物的AGL6同源基因可以准确的判断植物所属类群。通过蛋白预测说明pMADS4蛋白以同源或异源二聚体的形式存在,具有结合DNA的功能。
The MADS box gene pMADS4 was cloned from Petunia hybrida by RT-PCR and sequenced. The sequencing results showed that the obtained gene shares 100% homology with the published sequence. This gene is 910 bp in length and contains an open reading frame (ORF) encoding a protein of 253 amino acid residues. This gene has the structure of a typical plant MADS box gene. This gene has the same sequence as Zeamays ZAG3, Dendrocalamus latiflorus DlMADS18, The homologies of AGL6 in Arabidopsis thaliana were 64%, 64% and 61% respectively, indicating that pMADS4 is a homologue of AGL6 in petunia, suggesting that they have similar functions and both promote flowering and regulate flower Organ formation. Phylogenetic tree analysis showed that AP1 / AGL9 subfamilies were divided into three branches of SEP, AGL6 and AP1, and AGL6 branches were further divided into three groups, corresponding to gymnosperm, monocotyledon and dicotyledonous groups respectively. The predicted By isolating plants AGL6 homologous genes can accurately determine the plant belonging to taxa. Protein prediction shows that pMADS4 protein exists as homodimer or heterodimer, which has the function of DNA binding.