紫花苜蓿MsMYB33基因克隆、亚细胞定位及转录自激活检测

doi:10.11733/j.issn.1007-0435.2023.05.007

草地学报 ›› 2023, Vol. 31 ›› Issue (5): 1331-1337.DOI: 10.11733/j.issn.1007-0435.2023.05.007

紫花苜蓿MsMYB33基因克隆、亚细胞定位及转录自激活检测

王欣怡¹, 王晓倩², 刘亚玲³, 晁跃辉¹

1. 北京林业大学草业与草原学院, 北京 100083;
2. 北京泰德制药股份有限公司, 北京 100176;
3. 蒙草生态环境(集团)股份有限公司, 内蒙古呼和浩特 010010

收稿日期:2023-01-29 修回日期:2023-03-01 出版日期:2023-05-15 发布日期:2023-05-31
通讯作者: 晁跃辉,E-mail:chaoyuehui@bjfu.edu.cn
作者简介:王欣怡(1999-)，女，汉族，河北邢台人，硕士研究生，主要从事草地植物生物技术研究，E-mail:nqzxwxy@163.com
基金资助:
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

Cloning,Subcellular Localization and Autonomous Transcriptional Activation Testing of MsMYB33 Gene from Medicago sativa

WANG Xin-yi¹, WANG Xiao-qian², LIU Ya-ling³, CHAO Yue-hui¹

1. School of Grassland Science, Beijing Forestry University, Beijing 100083, China;
2. Beijing Tide Pharmaceutical Co., Ltd, Beijing 100176, China;
3. Inner Mongolia M-Grass Ecology And Environment(Group) Co., Ltd, Hohhot, Inner Mongolia 010010, China

Received:2023-01-29 Revised:2023-03-01 Online:2023-05-15 Published:2023-05-31

摘要/Abstract

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1 641 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。

关键词: 紫花苜蓿, MsMYB33基因, 核定位, 自激活活性

Abstract: 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.

Key words: Medicago sativa, MsMYB33 gene, Localization in nuclear, Self-activation activity

中图分类号:

S963.22+3.3

王欣怡, 王晓倩, 刘亚玲, 晁跃辉. 紫花苜蓿MsMYB33基因克隆、亚细胞定位及转录自激活检测[J]. 草地学报, 2023, 31(5): 1331-1337.

WANG Xin-yi, WANG Xiao-qian, LIU Ya-ling, CHAO Yue-hui. Cloning,Subcellular Localization and Autonomous Transcriptional Activation Testing of MsMYB33 Gene from Medicago sativa[J]. Acta Agrestia Sinica, 2023, 31(5): 1331-1337.

0
/ / 推荐

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://manu40.magtech.com.cn/Jweb_cdxb/CN/10.11733/j.issn.1007-0435.2023.05.007

https://manu40.magtech.com.cn/Jweb_cdxb/CN/Y2023/V31/I5/1331

参考文献

[1] KLEMPNAUER K H,GONDA T J,BISHOP J M. Nucleotide sequence of the retroviral leukemia gene v-myb and its cellular progenitor c-myb:the architecture of a transduced oncogene[J]. Cell,1982,31(2):453-463
[2] PAZ-ARES J,GHOSAL D,WIENAND U,et al. The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators[J]. The EMBO Journal,1987,6(12):3553-3558
[3] 梁玉镯,陈新娜,陈东亮,等. MYB转录因子调控植物花青素生物合成研究进展[J]. 江苏农业科学,2022,50(22):55-64
[4] 关淑艳,焦鹏,蒋振忠,等. MYB转录因子在植物非生物胁迫中的研究进展[J]. 吉林农业大学学报,2019,41(3):253-260
[5] OGATA K,KANEI I C,SASAKI M,et al. The cavity in the hydrophobic core of Myb DNA-binding domain is reserved for DNA recognition and trans-activation[J]. Nature Structural Biology,1996,3:178-187
[6] JIA L,CLEGG M T,JIANG T. Evolutionary dynamics of the DNA-binding domains in putative R2R3-MYB genes identified from rice subspecies indica and japonica genomes[J]. Plant Physiology,2004,134(2):575-585
[7] 牛义岭,姜秀明,许向阳. 植物转录因子MYB基因家族的研究进展[J]. 分子植物育种,2016,14(8):2050-2059
[8] 黎猛,陈跃,胡凤荣. miR159-GAMYB途径调控植物生长发育的研究进展[J]. 生物技术通报,2021,37(9):234-247
[9] ALLEN R S,LI J Y,STAHLE M I,et al. Genetic analysis reveals functional redundancy and the major target genes of the Arabidopsis miR159 family[J]. Proceedings of the National Academy of Sciences of the United States of America,2007,104(41):16371-16376
[10] GOCAL G W,SHELDON C C,GUBLER F,et al. GAMYB-like genes,flowering,and gibberellin signaling in Arabidopsis[J]. Plant Physiology,2001,127(4):1682-1693
[11] 张波,张杰,张颜,等. GA信号因子SlMYB33对番茄开花时间及果实大小的影响[J]. 西北农业学报,2020,29(12):1859-1866
[12] MAO X,JIA D,LI A,et al. Transgenic expression of TaMYB2A confers enhanced tolerance to multiple abiotic stresses in Arabidopsis[J]. Functional & Integrative Genomics,2011,11:445-465
[13] ZHANG L,ZHAO G,XIA C,et al. Overexpression of a wheat MYB transcription factor gene,TaMYB56-B,enhances tolerances to freezing and salt stresses in transgenic Arabidopsis[J]. Gene,2012,505:100-107
[14] PIECZYNSKI M,MARCZEWSKI W,HENNIG J,et al. Down-regulation of CBP 80 gene expression as a strategy to engineer a drought-tolerant potato[J]. Plant biotechnology journal,2013,11(4):459-469
[15] SHARMA N,SAHU P P,PRASAD A,et al. The Sw5a gene confers resistance to ToLCNDV and triggers an HR response after direct AC4 effector recognition[J]. Proceedings of the National Academy of Sciences of the United States of America,2021,118(33):e2101833118
[16] RUBIO S I,WEIGEL D. Coordination of flower maturation by a regulatory circuit of three microRNAs[J]. PLoS Genetics,2013,9(3):e1003374
[17] GUO C,JIANG Y,SHI M,et al. ABI5 acts downstream of miR159 to delay vegetative phase change in Arabidopsis[J]. New Phytologist,2021,231(1):339-350
[18] 任明辉,张雨蓬,许涛,等. 紫花苜蓿R2R3-MYB亚家族鉴定与干旱胁迫下的表达分析[J]. 草地学报,2023,31(4):972-983
[19] 李春艳,王曦,周胜花,等. 白羊草R2R3-MYB转录因子的挖掘及对干旱胁迫反应的研究[J]. 草地学报,2020,28(6):1784-1790
[20] 严莉,王翠平,陈建伟,等. 基于转录组信息的黑果枸杞MYB转录因子家族分析[J]. 中国农业科学,2017,50(20):3991-4002
[21] 邢铮. 马铃薯基因StR2R3-MYB1的克隆与功能分析[D]. 长沙:湖南农业大学,2014:33-34
[22] 靳亚忠,齐娟,何淑平,等. R2R3-MYB转录因子调控果蔬品质形成的研究进展[J]. 农业生物技术学报,2021,29(2):364-374
[23] 吕欣,尹文,雷迎峰,等. HCV截短型E2蛋白在酵母双杂交系统中的表达及自激活作用检测[J]. 生物技术,2008(5):9-11
[24] 董笛,王焕英,滕珂,等. 日本结缕草ZjHCT基因的克隆及转录自激活检测[J]. 分子植物育种,2019,17(14):4617-4622
[25] 丁恺,麻鹏达,贾彦彦,等. 丹参3个R2R3-MYB转录因子的亚细胞定位和自激活活性分析[J]. 西北农业学报,2018,27(4):586-594

紫花苜蓿MsMYB33基因克隆、亚细胞定位及转录自激活检测

Cloning,Subcellular Localization and Autonomous Transcriptional Activation Testing of MsMYB33 Gene from Medicago sativa

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	陈超超, 李贺洋, 闫雨欣, 宋晴, 黄悠, 热孜亚·艾力, 张铁军. 20份紫花苜蓿种质对黄萎病的抗性评价[J]. 草地学报, 2023, 31(5): 1425-1434.
[2]	路文杰, 吴聪, 齐晋云, 景亚泓. 生物炭和接种剂配施对紫花苜蓿生物量和土壤特征的影响[J]. 草地学报, 2023, 31(5): 1578-1587.
[3]	任明辉, 张雨蓬, 许涛, 朱慧森, 岑慧芳. 紫花苜蓿R2R3-MYB亚家族鉴定与干旱胁迫下的表达分析[J]. 草地学报, 2023, 31(4): 972-983.
[4]	谭璐娜, 彭嘉文, 许环宇, 张瀚文, 夏厚胤, 苗愉祺, 崔祎琳, 苑旺, 马志超, 同雨涛, 高景慧. 紫花苜蓿胚性愈伤组织诱导与增殖的研究[J]. 草地学报, 2023, 31(4): 1057-1063.
[5]	刘育书, 曹岭, 余舟昌, 田兴芳, 孙琳, 时嘉翊, 张伟, 谢治国, 雷秀云, 俞靓, 杨培志. 紫花苜蓿生物量分配对植物-土壤化学计量的响应[J]. 草地学报, 2023, 31(4): 1143-1153.
[6]	谭瑶, 乔浪, 李玲, 杨佳乐, 韩海斌, 朱猛蒙, 张志强, 倪鹏. 不同诱虫色板及悬挂方式对紫花苜蓿田牧草盲蝽的诱集效果[J]. 草地学报, 2023, 31(4): 1226-1233.
[7]	孙林, 肖燕子, 都帅, 张福金, 刘广华, 张欣昕, 乌尼尔, 娜娜, 赵熠, 薛艳林. 贮藏时间与蒙药黄芩对紫花苜蓿草颗粒维生素和化学成分的影响[J]. 草地学报, 2023, 31(4): 1242-1253.
[8]	冯启贤, 张磊, 李妍, 凌文卿, 李珏, 周燚, 阳伏林, 周晶. 甲酸处理对苜蓿青贮品质及瘤胃降解率的影响[J]. 草地学报, 2023, 31(4): 1264-1272.
[9]	苏立娜, 麻冬梅, 李嘉文, 张玲, 侯汶君. 外源褪黑素对盐胁迫下两种紫花苜蓿生理及光合特性的影响[J]. 草地学报, 2023, 31(3): 726-732.
[10]	刘琳絮, 王铁梅, 张霁, 苏德荣. 紫花苜蓿根系主要功能性状对分根区交替灌溉的响应[J]. 草地学报, 2023, 31(3): 852-859.
[11]	侯燕红, 生小燕, 韩博, 姜华, 何承刚. 干旱和高温胁迫下紫花苜蓿PEPC酶活性变化及其基因序列分析[J]. 草地学报, 2023, 31(2): 330-336.
[12]	刘艳君, 胡桂馨, 温雅洁, 周生英. 施K对苜蓿N，P，K吸收利用及其抗蓟马的影响[J]. 草地学报, 2023, 31(2): 418-425.
[13]	张宏, 黄荣, 姚博, 张玉娟, 张振粉. 桃色欧文氏菌Cp2脂多糖对紫花苜蓿幼苗生长的影响[J]. 草地学报, 2023, 31(2): 615-622.
[14]	徐小博, 王鑫尧, 郁伟杰, 苗一凡, 徐博. 紫花苜蓿MsJMT基因的克隆及其与育性的关联分析[J]. 草地学报, 2022, 30(9): 2306-2314.
[15]	孟翔, 孙伶俐, 谢开云, 刘伟, 褚皓清, 赵越. 灌溉、施磷量及AM真菌对紫花苜蓿产量和水磷利用效率的影响[J]. 草地学报, 2022, 30(9): 2356-2364.