蒺藜苜蓿MtROP10基因的克隆及其功能初探

Cloning of MtROP10 in Medicago truncatula and Its Functional Study

目的:克隆并研究蒺藜苜蓿ROP基因的功能,为研究该基因家族在共生途径中的作用提供依据。方法:采用RACE方法,从蒺藜苜蓿中克隆MtROP基因,利用生物信息软件比对同源性及ROP蛋白特征结构分析,利用RT-PCR方法分析该基因的组织特异性表达,构建该基因的过表达载体并转化拟南芥。结果:获得了蒺藜苜蓿ROP家族中与拟南芥ROP10高度同源的MtROP10全长序列。氨基酸编码序列具有明显的ROP家族蛋白的结构域特征。该基因在花中高表达,根中低表达。拟南芥中过表达MtROP10,可导致根毛变粗、变短、分叉。结论:MtROP10属于植物ROP家族蛋白,可能在根毛的极性生长方面具有较为重要的功能。

Objective：The objective of this study was to clone Medicago trunctula ROP gene and study its function.Method：the full-length Medicago trunctula ROP gene was cloned with RACE method according to the EST sequences.The function domain of the gene was predicted by bioinformatics software.The tissue expression level of the gene was detected by RT-PCR.Over-expression of MtROP10 vector was Constructed and transformed into Arabidopsis.Result：The full-length sequence was obtained by 3＇RACE.Analysis result shown that encoded sequence had the characteristics domain of ROP family and shared high similarity with AtROP10 in Arabidopsis.RT-PCR showed that expression level is difference in different tissues.Over-expression of MtROP10 gene in Arabidopsis resulted in swelling,shortening and branching root hair.Conclusion：MtROP10 is a member of plant ROP family,and probably plays a vital role in the polar growth of root hair.