论文部分内容阅读
采集森林草莓(Fragaria vesca)植株叶片,TRIzol法提取样本总RNA,设计特异性引物RT-PCR扩增转录因子MYB 12基因,获得1条约1 kb大小的特异性片段,克隆并测序。序列分析表明,该片段序列全长988 nts,含有完整的MYB 12基因开放阅读框(ORF),ORF长度为855 nts,编码285个氨基酸。序列比对发现,森林草莓MYB 12基因与来源于5种蔷薇科植物MYB基因的序列相似性较高,为75%~76%,而与其他7种非蔷薇科植物MYB基因的序列相似性相对较低,为47%~68%。构建MYB基因系统关系树,发现森林草莓与其他5种蔷薇科植物的MYB基因聚成一个分支,说明森林草莓与蔷薇科植物亲缘关系较近。生物信息学分析表明,MYB 12蛋白具有一定的亲水性,其N-端有2个MYB结构域,属于R2R3-MYB亚族。MYB 12蛋白二级结构主要以α-螺旋为主,并含有一些不规则的卷曲(Random coil)结构。这为研究该基因在植物生长发育及生理代谢过程中的调控机制奠定了基础。
The leaves of Fragaria vesca plants were harvested and the total RNA was extracted by TRIzol method. The MYB 12 gene was amplified by RT-PCR and cloned and sequenced. Sequence analysis showed that the full length of this fragment was 988 nts, which contained the complete open reading frame (ORF) of MYB 12 gene. The ORF length was 855 nts and encoded 285 amino acids. The sequence alignment showed that the MYB 12 gene in forest strawberry had a high similarity (75% -76%) with the MYB gene from five species of Rosaceae, but had similar sequence similarity with MYB gene in other seven non-Rosaceae plants Lower, 47% to 68%. The MYB gene phylogenetic tree was constructed and found to be a branch of the MYB gene between forest strawberry and other five species of Rosaceae, indicating that the forest strawberry is more closely related to the Rosaceae plants. Bioinformatics analysis showed that the MYB 12 protein has a certain hydrophilicity and has 2 MYB domains at the N-terminus, belonging to the R2R3-MYB subfamily. The secondary structure of MYB 12 protein is dominated by α-helix and contains some irregular random coil structures. This laid the foundation for studying the regulatory mechanism of this gene in plant growth and development and physiological metabolism.