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森林草莓转录因子MYB 12基因克隆及蛋白特性分析
陈静,胡亚会,张享享,左登攀,江彤
0
(安徽农业大学植物保护学院,合肥 230036)
摘要:
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。
关键词:  森林草莓  MYB 12基因  克隆  蛋白特性分析
DOI:10.13610/j.cnki.1672-352x.20150302.007
投稿时间:2014-12-15
基金项目:国家公益性行业(农业)科研专项(201303028)资助。
Cloning and protein property analysis of the transcription?factor MYB 12 gene in woodland strawberry (Fragaria vesca)
CHEN Jing,HU Yahui,ZHANG Xiangxiang,ZUO Dengpan,JIANG Tong
(School of Plant Protection, Anhui Agricultural University, Hefei 230036)
Abstract:
Total RNA was extracted from leaves of woodland strawberry (Fragaria vesca) using TRIzol method. A specific primer pair was designed to amplify the MYB 12 gene encoding a transcription factor by RT-PCR. A specific fragment about 1 kb was amplified and then cloned and sequenced. Sequence analysis showed that the fragment consisted of 988 nucleotides (nts), including a complete 855 nts ORF encoding 285 amino acids. Sequence comparison revealed that MYB 12 of woodland strawberry shared a relatively high sequence similarity (75%-76%) with that of five other Rosaceae plants and a relatively low sequence similarity (47%-68%) with that of seven non-Rosaceae plants. A phylogenetic tree based on the nucleotide sequence alignment of the MYB genes was constructed. The MYB gene of woodland strawberry and five other Rosaceae plants were clustered into the same branch, which indicated that woodland strawberry had a close relationship with Rosaceae plants. Bioinformatics analyses demonstrated that MYB 12 protein was hydrophobic. Its N-terminal contained two MYB structural domains belonging to subtribe R2R3-MYB. The secondary structure of MYB 12 protein was dominated by α-helix and some random coils were included. This research will provide a basis for identifying the role of the MYB gene in regulating plant growth and development, as well as physiological and metabolic processes.
Key words:  woodland strawberry (Fragaria vesca)   MYB 12 gene  cloning  protein property analysis

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