蒺藜苜蓿(Medicago truncatula)MtAP2转录因子的克隆及自激活检测

Cloning and Self-activation Detection of MtAP2 Transcription Factor in Medicago truncatula

AP2转录因子在植物生长发育过程中具有重要作用,为深入研究蒺藜苜蓿MtAP2转录因子的功能,于蒺藜苜蓿中克隆MtAP2转录因子,进行生物信息学分析,构建诱饵表达载体PGBKT7-MtAP2并转入酵母菌中,进行自激活活性检测。结果显示,本研究成功克隆了蒺藜苜蓿MtAP2转录因子,其开放阅读框为624 bp,编码207个氨基酸。生物信息学分析显示,蒺藜苜蓿MtAP2蛋白与鹰嘴豆的亲缘关系最为接近。诱饵表达载体PGBKT7-AP2构建成功,转入到Y2H酵母菌中,且通过在营养缺陷型培养基培养证明其没有自激活活性。本研究成功克隆了蒺藜苜蓿MtAP2转录因子,构建了酵母诱饵表达载体PGBKT7-MtAP2,并证明其不存在自激活现象,后续可进行互作蛋白筛选。

AP2 transcription factor plays an important role in plant growth and development.To further study the function of MtAP2 transcription factor in Medicago truncatula,MtAP2 transcription factor in Medicago truncatula was cloned,bioinformatics analysis was conducted,the bait expression vector PGBKT7-MtAP2 was constructed and transferred to yeast,and self-activating activity was detected.The results showed that the MtAP2 transcription factor of Medicago truncatula was successfully cloned in this study,with an open reading frame of 624 bp and encoding 207 amino acids.Bioinformatics analysis showed that Medicago truncatula MtAP2 protein was most closely related to Cicer arietinum.The bait expression vector PGBKT7-AP2 was successfully constructed,transferred into Y2H yeast,and proved to have no self-activating activity by culturing in an auxotrophic medium.In this study,the MtAP2 transcription factor of Medicago truncatula was successfully cloned,and the yeast bait expression vector PGBKT7-MtAP2 was constructed.It was proved that there was no self-activation phenomenon,and subsequent interaction protein screening can be performed.