论文部分内容阅读
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。
Epi BL was sprayed on Dongnong 42 soybean to determine the effect of brassinolide (BR) on the expression of GmRAV gene. Meanwhile, epi BL was exogenously added to Tm generation GmRAVox tobacco to study the effect of GmRAV Whether or not the gene is also involved in the BR signal inhibits plant growth and development. The results showed that BR inhibited the expression of GmRAV gene and epiBL did not promote the stem elongation of GmRAVox tobacco plants, indicating that GmRAV may be a negative regulator of BR signaling pathway to promote plant growth and development. Meanwhile, the cloned GmRAV gene in soybean was cloned into the prokaryotic expression vector pET28a to construct a recombinant protein pET28a-GmRAV plasmid fused with His-tag, which was then transformed into E. coli BL21 (DE3) Under different temperatures, 0.5 mmol·L -1 IPTG was induced with a induction time of 6 h. The result of SDS-PAGE showed that the recombinant protein with the molecular weight of about 40 kDa was expressed at different temperatures, and the low temperature could obviously enhance the solubility of GmRAV protein.