梭梭NAC转录因子家族基因的克隆和转录激活分析

Cloning and Transcriptional Activation Analysis of NAC Gene Family from Haloxylon ammodendron

以梭梭NAC转录组测序结果设计特异引物,成功克隆5个梭梭NAC转录因子。通过与其它物种NAC蛋白进行多重比对,表明克隆的5条序列具有NAC转录因子所共有的5个典型亚结构域。通过构建p GBKT7融合表达载体,将其转入AH109酵母菌中进行转录激活功能分析。结果显示含有Ha NAC10、Ha NAC17、Ha NAC18、Ha NAC21转录因子的重组酵母菌可以在SD-Trp-His-Ade缺陷培养基平板上生长,X-gal检测显蓝色。这表明Ha NAC10、Ha NAC17、Ha NAC18、Ha NAC21可以激活下游报告基因表达,说明其具有转录激活活性;Ha NAC16不具备转录激活活性。为进一步分析梭梭NAC转录因子功能和调控机制提供帮助。

Five Haloxylon ammodendron NAC transcription factors were successfully cloned with primers designing by Haloxylon transcriptome sequencing. It was indicated that the five cloned sequence had five typical sub-domains, through multiple alignment. By building pGBKT7 fusion expression vector and being carried into AH109 yeast, transcriptional activation was analyzed. The results showed that the recombinant yeast containing HaNACIO, HaNAC17, HaNAC18, HaNAC21 transcription factors could grow on SD-Trp-His-Ade defective medium, X-gal detection showed blue. This indicated that HaNACIO, HaNAC17, HaNAC18, HaNAC21 could activate the expression of downstream reporter gene, and had transcriptional activation activity; HaNA C16 do not have transcriptional activation activity. This experiment could be helpful for the further analysis of its function and regulation mechanism.