The Expression,Purification And Preliminary Bioinformatics Analysis Research Of Sex Determination Factors

Sex determination refers to the process that results in gender differentiation,and formats the group differences of male and female populations in the eukaryotic life.These sex determination signals are closely related to the conservative molecular mechanism,which initiates the initial development of the female or male.Sex determination mechanisms are diverse,involving the evolution of different taxa,evolving a variety of sex determination systems.Sex determination system has a long history,in the process of higher biological evolution,different species used in different ways to determine the gender.However even with the same sex chromosome group,the sex determination factors of different animals are not the same.At present,there are just a few gender determinants identified.Besides to the mammalian sex determination gene Sry,in recent years,sex determination gene dmrt1 bY of medaka,silkworm Fem piRNA,Aedes albopictus nix,As well as transcription factors FOXL2/FOXL3 which play an important role in the organism's gender maintenance mechanism and estrogen receptor EsR2 are also found.Through a series of gene knockout experiments and functional validation experiments,people gradually improved the gender control network,and proposed three possible modes to determine the path of gender.However,they are unknown about how does transcription factors recognize their target genes specifically,how they interact,how to maintain dose balance,whether there are other functional proteins involved in the pathway,how piRNA combine to the chaperone and how the complex cuts the target Genes.But the problems involving in the level of molecular,can only rely on structural biology method to solve by analyzing the transcription factor and chaperone protein structure,which is currently best way to perfect these signal paths and to clarify the molecular mechanism of gender determination precisely.In this study,we respectively expressed and purified the two genes of medaka successfully,which were the key factors of sex determination DMY/dmrt1 bY and the female sex maintenance transcription factor FOXL3.Through the continuous adjustment of the purification program,to a certain extent,the state of recombinant protein was improved.As an important first step in structural biology research,this work laid the foundation for the subsequent exploration of crystal optimization conditions.