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《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:

第46卷

期数:

2018年第23期

页码:

50-55

栏目:

生物技术

出版日期:

2018-12-05

文章信息/Info

Title:

Cloning and prokaryotic expression of MYB transcription factor PcMYB2 gene from Polygonum cuspidatum

作者:

李晓筱;  徐俊雄;  刘婵;  王岩岩;  伍翔;  覃建兵;  柳忠玉

长江大学生命科学学院，湖北荆州 434025

Author(s):

Li Xiaoxiao; et al

关键词:

虎杖; MYB转录因子; PcMYB2; 原核表达; 苯丙烷代谢; 原花青素

Keywords:

-

分类号:

S567.23⁺9.01

DOI:

-

文献标志码:

A

摘要:

为了探究MYB(髓细胞组织增生病毒癌基因同源物)转录因子在虎杖苯丙烷代谢途径中的作用，以虎杖叶片为材料，通过cDNA末端快速扩增(RACE)技术获得1个MYB转录因子，命名为PcMYB2，GenBank登录号为MG020557。序列分析表明，PcMYB2基因的cDNA序列全长为 938 bp，开放阅读框(ORF)为738 bp，编码245个氨基酸，推测蛋白质分子质量为27.917 ku。PcMYB2编码的氨基酸序列具有R2R3-MYB类转录因子的共同特征，其N端具有R2、R3这2个MYB结构域，并且R3结构域中包含1个能与bHLH转录因子相互作用的［D/E］Lx₂［R/K］x₃Lx₆Lx₃R基序。进化树分析表明，PcMYB2蛋白与苜蓿中参与原花青素调控的MtPAR聚为1组。将该基因的ORF片段连接到原核表达载体pET20b中，构建融合表达载体pET20b-PcMYB2，转化到大肠杆菌(Escherichia coli) Rosetta(DE3)中进行表达。SDS-PAGE(十二烷基硫酸钠-聚丙烯酰胺凝胶电泳)检测显示表达蛋白与预期大小一致，表明成功克隆PcMYB2基因的cDNA全长序列，构建的原核表达载体pET20b-PcMYB2可用于该基因的功能研究。

Abstract:

-

参考文献/References:

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备注/Memo

备注/Memo:

收稿日期：2018-03-25
基金项目：国家自然科学基金(编号：81803670)；国家级大学生创新项目(编号：201810489013)；长江大学校级大学生创新训练计划(编号：2017073)。
作者简介：李晓筱(1998—)，女，湖北仙桃人，主要从事药用植物次生代谢研究。E-mail：xufzy2016@126.com。
通信作者：柳忠玉，博士，讲师，主要从事中药生物技术研究。E-mail：zyliu2004@126.com。

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更新日期/Last Update: 2018-12-05