Construction Toxicity Detection and Autoactivation of a Yeast Two-hybrid Bait Vector Containing a Sisal AhMADS8937 Gene
MADS-box is an important transcription regulator in plants,which is involved in seed dormancy and germination,root growth,flower organ development,fruit maturation and other vegetative and reproductive growth processes.The results of previous studies indicated that A hMA DS8937 may be involved in regulating the development of bulbil of sisal.In view of the fact that MADS-box protein can regulate plant development in the form of complex,in order to screen the proteins interacting with AhMADS8937 protein in sisal(Agave hybrid No.11648),the bait of pGBKT7-AhMADS8937 vector was constructed.The restriction enzyme analysis and DNA sequencing showed that the vector was successfully constructed.The recombinant vector was transformed into Y2HGold yeast to detect its toxicity and autoactivation.The result showed that the bait vector had no toxicity to Y2HGold.However,it had autoactivation,which was inhibited by 5 mmol/L 3-AT.The study successfully constructed the bait vector of AhMADS8937 for yeast two-hybrid system,which laid a foundation for screening the proteins interacting with AhMADS8937.
Agave hybrid No.11648AhMADS8937Yeast two-hybridBait vectorToxicity and autoactivation
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