摘要
MYB(V-myb Avian myeloblastosis viral Oncogene Homolog)转录因子数量较多、功能多样,在植物的发育过程中起着十分重要的作用。为了探究紫花苜蓿(Medicago sativa)中MsMYB33基因的亚细胞定位及自激活特性,本研究从紫花苜蓿中克隆MsMYB33基因,该基因开放阅读框为1641 bp,编码多肽含有546个氨基酸。遗传进化树显示MsMYB33蛋白与鹰嘴豆(Cicer arietinum)中的CaMYB33蛋白同源关系最高。将MsMYB33与黄色荧光蛋白(Yellow fluorescent protein,YFP)进行融合表达,结果显示:MsMYB33蛋白定位于细胞核。通过DNA重组技术成功构建pGBKT7-MYB33酵母表达载体,并转化到Y2HGold酵母菌中。酵母自激活检测结果显示,MsMYB33具有自激活活性,进一步分析发现激活区域位于C端。本研究为进一步研究紫花苜蓿MsMYB33基因及MYB转录因子家族提供了科学依据。
MYB(V-myb Avian myeloblastosis viral Oncogene Homolog)transcription factors,with numerous members and diverse functions,play important roles in the developmental processes of plants.To investigate the subcellular localization and self-activation properties of the MsMYB33 gene in Medicago sativa,the MsMYB33 gene was cloned from M.sativa,which had an open reading frame of 1,641 bp and encoded a polypeptide with 546 amino acids.Upon the phylogenetic analysis,that MsMYB33 protein was most closely related to the CaMYB33 protein in Cicer arietinum.MsMYB33 was fused with yellow fluorescent protein(YFP)for expression,and the results showed that the MsMYB33 protein was localized in the nucleus.A pGBKT7-MYB33 yeast expression vector was successfully constructed by DNA recombination technology and transformed into the Y2HGold yeast strain.Yeast self-activation assays showed that MsMYB33 had a self-activation activity,and from the further analysis that the self-activation domain of MsMYB33 was located at the C-terminus.This study provided a scientific basis for further research on the MsMYB33 gene and the MYB transcription factors family in M.sativa.
作者
王欣怡
王晓倩
刘亚玲
晁跃辉
WANG Xin-yi;WANG Xiao-qian;LIU Ya-ling;CHAO Yue-hui(School of Grassland Science,Beijing Forestry University,Beijing 100083,China;Beijing Tide Pharmaceutical Co.,Ltd,Beijing 100176,China;Inner Mongolia M-Grass Ecology And Environment(Group)Co.,Ltd,Hohhot,Inner Mongolia 010010,China)
出处
《草地学报》
CAS
CSCD
北大核心
2023年第5期1331-1337,共7页
Acta Agrestia Sinica
基金
内蒙古自治区科技厅内蒙古自治区科技重大专项项目专题“抗寒高产苜蓿新品种培育”(2022 JBGS00160302)资助。
关键词
紫花苜蓿
MsMYB33基因
核定位
自激活活性
Medicago sativa
MsMYB33 gene
Localization in nuclear
Self-activation activity
