大豆GmRAV基因参与BR信号传导途径及重组蛋白可溶性分析  

Soybean GmRAV Gene Involved in BR Signaling Pathway and Analysis on Recombinant Protein Solubility

作　　者：杨春亮[1] 孙浩航 张涵朔 赵琳[2] YANG Chun-liang SUN Hao-hang ZHANG Han-shuo ZHAO Lin(Basic Medical Science School, Harbin Medical University, Harbin 150081, China 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)

机构地区：[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

出　　处：《大豆科学》2016年第6期928-931,957,共5页Soybean Science

基　　金：黑龙江省教育厅面上项目(12541465)

摘　　要：对大豆东农42喷施表油菜素内酯(epi BL),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epi BL,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p ET28a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。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 inhib- it 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.

关 键 词：大豆 BR GmRAV蛋白 原核表达 可溶性分析 

分 类 号：S565.1[农业科学—作物学]