论文部分内容阅读
课题组前期对干旱胁迫下大豆转录组的数据分析发现大豆Glyma.13G115900基因编码一个RING/U-box蛋白,其表达水平受干旱胁迫影响显著。本研究以垦丰16大豆为试验材料,克隆Glyma.13G115900基因。氨基酸多重序列比对表明其编码的蛋白与其它物种都具有高度保守的RING/U-box结构域。构建原核表达载体pET-29b-Glyma.13G115900转化到大肠杆菌中,Glyma.13G115900蛋白在大肠杆菌中能够表达。荧光定量PCR分析表明Glyma.13G115900基因的表达量受PEG、NaCl和ABA的影响显著,但基本不受冷胁迫的诱导。经PEG和NaCl处理后,该基因表达量与CK相比呈现出显著下降的趋势,PEG处理的表达量变化比NaCl下调的更明显;在ABA诱导下与CK相比该基因的mRNA丰度呈现出先上升后下降的趋势,在4 h表达量出现峰值,推测该基因可能通过依赖于ABA途径参与非生物胁迫应答。以上结果为进一步深入研究该基因的调控途径奠定基础。
Our previous data analysis of the soybean transcriptome under drought stress found that soybean Glyma.13G115900 gene encodes a RING / U-box protein whose expression level is significantly affected by drought stress. In this study, Kenfeng 16 soybean as experimental materials, cloning Glyma.13G115900 gene. Multiple sequence alignment of amino acids indicates that the protein encoded by it has a highly conserved RING / U-box domain with other species. Construction of prokaryotic expression vector pET-29b-Glyma.13G115900 transformed into E. coli, Glyma.13G115900 protein can be expressed in E. coli. Fluorescent quantitative PCR analysis showed that the expression of Glyma.13G115900 gene was significantly affected by PEG, NaCl and ABA, but not by the cold stress. After treatment with PEG and NaCl, the expression of this gene showed a significant decrease compared with that of CK, and the change of PEG treatment was more pronounced than that of NaCl. Compared with CK, ABA-induced mRNA abundance At first, the expression peak appeared at 4 h, suggesting that this gene may participate in abiotic stress response by relying on ABA pathway. The above results lay a foundation for further study on the regulation of this gene.