刚毛柽柳ThbHLH1转录因子识别的顺式作用元件的鉴定

Identification of Cis-acting Elements Recognized by ThbHLH1 Transcription Factor of Tamarix hispida

摘要:
目的本研究拟对ThbHLH1所识别的顺式作用元件进行鉴定，进一步揭示ThbHLH1调控抗逆基因表达的机理。
方法利用以转录因子为中心的酵母单杂交系统鉴定ThbHLH1所识别的顺式作用元件；将元件与报告基因融合构建报告载体（pCAM-Cis），通过基因枪法将报告载体与效应载体35S：ThbHLH1共转化烟草叶片，在盐、干旱胁迫下比较GUS酶活。
结果鉴定出2段能够与ThbHLH1转录因子结合的DNA序列：分别为CCGAAA（LTRE1）和TGAC（WRKY710S）。
结论在盐或干旱胁迫下，ThbHLH1通过与LTRE1或WRKY710S元件作用来激活基因表达。

Abstract:
Objective To further study the mechanism of ThbHLH1 overexpression on improving the salt and drought tolerance of Tamarix hispida.
Method The cis-acting elements recognized by ThbHLH1 were identified with the transcription factor-centered yeast one hybridization (TF-centered Y1H) method. To confirm the results of TF-centered Y1H, the effector construct 35S:ThbHLH1 was co-transformed with each reporter constructs pCAM-Cis into tobacco leaves with the particle bombardment method, and then the GUS activity was determined under salt or drought stress.
Result ThbHLH1 could recognize LTRE1 (CCGAAA) and WRKY710S (TGAC) Cis-acting elements.
Conclusion The results show that ThbHLH1 activates gene expression under salt or drought stress by interacting with LTRE1 or WRKY710S motifs.