Molecular Mechanism Of A Zinc Finger Protein MsZFN From Alfalfa

As"the king of forages",Medicago sativa is the earliest cultivation,the most extensive distribution and the most important perennial quality legumes which has high yield, good palatability and rich protein,but the salt tolerance limited its planting area. In these years,improve plant cultivar by genetic engineering technology become a hot spot of biological engineering research.Transcription factors play important role in the regulation of plant growth and development,as well as its response to environment. Zinc finger protein is designateed as it contains finger-like domain that has Zn²⁺ binding ability. As superfamily with typical zinc finger domains,zinc finger proteins have involved in any growth and development aspects of organism and have been widely found in many animals and plants. They have been classified into nine types according to the structural and functional diversities.The CCCH-type zinc finger is less common compared to other zinc fingers and represents approximately 0.8% of all zinc fingers.A novel CCCH-type zinc finger gene, Mszfn,which was screened from the suppression subtraction hybridization (SSH) cDNA library that induced by salt stress. The provious research show that the exact function of the protein is unknown, but it may play an important role in organisms.It's necessary to take the study of molecular mechanism of MsZFN protein.This work aims to search for the proteins interacting with MsZFN and study the mechanism of MsZFN through their relationship .In this work, yeast two-hybrid system"Matchmaker? Library Construction & Screening Kits"was utilized to screen interactor proteins with MsZFN from Alfalfa cDNA library. Several proteins showed interaction to MsZFN,two protein was gave the main research,one is MsZFN,and another resembled to cyanase,named it MZ69C.Bimolecular fluorescence complementation (BIFC) used for the further authentication. The result also showed MsZFN interact to MsZFN and MZ69C.According to finger position,the deletion mutants of MsZFN were construct as bait vector. Interaction regions analysis of MsZFN showed that have inhibitor beteen the 36aa to the 81aa when it interacted with MZ69C.