[1]杨春亮,孙浩航,张涵朔,等.大豆GmRAV基因参与BR信号传导途径及重组蛋白可溶性分析[J].大豆科学,2016,35(06):928-931.[doi:10.11861/j.issn.1000-9841.2016.06.0928]
　YANG Chun-liang,SUN Hao-hang,ZHANG Han-shuo,et al.Soybean GmRAV Gene Involved in BR Signaling Pathway and Analysis on Recombinant Protein Solubility[J].Soybean Science,2016,35(06):928-931.[doi:10.11861/j.issn.1000-9841.2016.06.0928]
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大豆GmRAV基因参与BR信号传导途径及重组蛋白可溶性分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年06期 页码: 928-931 栏目: 出版日期: 2016-11-20
Title:
Soybean GmRAV Gene Involved in BR Signaling Pathway and Analysis on Recombinant Protein Solubility
作者:
杨春亮1; 孙浩航1; 张涵朔1; 赵琳2
1.哈尔滨医科大学 基础医学院，黑龙江 哈尔滨150081; 2.东北农业大学 大豆生物学教育部重点实验室/农业部东北大豆生物学与遗传育种重点实验室，黑龙江 哈尔滨150030
Author(s):
YANG Chun-liang1;  SUN Hao-hang1;  ZHANG Han-shuo1;  ZHAO Lin2
（1.Basic Medical Science School, Harbin Medical University, Harbin 150081, China; 2.Key Laboratory of Soybean Biology in Chinese Ministry of Education Northeast Agricultural University/Key Laboratory of Soybean Biology and Breeding (Genetics) of Chinese Agriculture Ministry，Harbin 150030, China）
关键词:
大豆;  BR; GmRAV蛋白; 原核表达;  可溶性分析
Keywords:
Soybean; GmRAV protein;  Prokaryotic expression;  Solubility analysis
DOI:
10.11861/j.issn.1000-9841.2016.06.0928
文献标志码:
A
摘要:
对大豆东农42喷施表油菜素内酯（epiBL），进行GmRAV定量，研究油菜素内酯(BR)对GmRAV基因表达的影响，同时对T5代GmRAVox烟草外源添加epiBL，研究GmRAV基因是否也参与BR信号从而抑制植物的生长发育。结果表明：BR抑制GmRAV基因表达，并且外源添加epiBL并未促进GmRAVox烟草植株茎延伸，表明GmRAV可能为BR促进植物的生长发育信号传导途径的负调节因子。同时，将大豆中已克隆的GmRAV基因克隆到原核表达载体pET28a上，构建与His标签融合的重组蛋白pET28a-GmRAV质粒，转化大肠杆菌BL21(DE3) 中，对其进行37℃、28℃两种不同温度下0.5mmol·L-1 IPTG 诱导，诱导时间为6 h。SDS-PAGE 电泳结果表明: 分子量大约为40 kDa的重组蛋白在不同温度下均表达，低温可以明显增强 GmRAV蛋白的可溶性。
Abstract:
Soybean DN42 was sprayed by epiBL to study the effect of BR on GmRAV expression by real-time quantative PCR Meantime, T5 generation GmRAVox tobaccos were treated with epiBL to study if GmRAV was involved in BR signaling to inhibit the plant growth and development.The results showed that the GmRAV mRNA expression level was inhibited by BR, and the exogenous epiBL addition did not promote GmRAVox tobacco stem elongation，which indicated that GmRAV might be a negative regulator in BR signaling pathway. Furthermore, soybean GmRAV was constructed into prokaryotic expression vector pET28a and the recombinant plasmid pET28a-GmRAV was transformed into E.coli BL21 (DE3) to get the expression protein for further study． The bacterial was induced by 0.5mmol·L-1?IPTG in two different temperature 37℃ and 28℃. The result indicated that an 40 kDa recombinant protein was all expressed with the treatment of different temperature by SDS-PAGE．The lower temperature could increase protein solubility．

参考文献/References:

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备注/Memo

基金项目：黑龙江省教育厅面上项目（12541465）。

第一作者简介：杨春亮(1980-) ，男，硕士，副研究员，主要从事大豆生物技术研究工作。E-mail: 360473662@qq.com。
通讯作者：赵琳（1980-），女，研究员，博导，主要从事大豆生物技术研究。E-mail:zhaolinneau@126.com。

更新日期/Last Update: 2016-12-09