Regulatory Role Of SRF In Restarting Diapause Embryos Of Artemia Sinica

Artemia sinica is a crustacean living in a high-salt environment.Under adverse conditions,Artemia will appear diapause.When the environmental conditions are favorable to the development of gastrointestinal embryos,the development of Artemia sinica.Serum response factor(SRF)is a highly conserved cis-acting factor and one of the members of the MADS transcription factor superfamily.SRF can bind to regulatory elements in many gene promoters.SRF promotes the migration of mesoderm cells,plays an important role in the extension of the embryonic body axis,and is essential for embryonic development.The protein can regulate the activity of many early genes(such as c-fos),thereby participating in the regulation of cell cycle,apoptosis,growth and differentiation.However,the mechanism of SRF in the removal of diapause is still unclear.In this study,the full-length c DNA sequence of As-srf was cloned by RACE technology.The full-length sequence of As-srf is 1223 bp,including the open reading frame 912 bp,149bp3'-UTR and 162 bp 5'UTR,co-coding 303 amino acids,the molecular weight of the protein is about 34 KDa,the protein is a hydrophilic protein,the isoelectric point is 6.32;immunofluorescence experiments show that As-SRF is expressed in all parts of the Chinese Artemia,indicating that this protein has no tissue and Organ specificity;real-time fluorescence quantitative q PCR and Western Blot results proved that the expression levels of As-srf and As-SRF are basically the same,the expression level increases from 0 to 20 h after development,and the expression level gradually decreases after 20 h,indicating that the serum Response factors may play an important role in embryo restart and early embryo development.In response to temperature and salinity stress,the expression of As-SRF showed a gradual downward trend as the temperature decreased and salinity increased,indicating that As-SRF played a role in the response to low temperature and high salt stress.Important regulation effect;at the same time,Western bloting technology was used to detect the expression levels of As-JUN,As-C-FOS,As-SRE and As-ELK1.The results showed that the expression trend of each protein was similar to As-SRF,and it was speculated This protein can form AP-1complex and mediate the transformation of G1 phase to S phase in the cell cycle;using small RNA interference technology,knocking down the m RNA expression level of As-srf,Artemia sinica appear to be slow to develop and deform Abnormal conditions.The above research results show that As-SRF played an important role in the restart of diapause embryos and early embryo development in Artemia sinica.In this paper,through the study of As-SRF in the early embryonic development of Artemia sinica and the restart of diapause embryos,it is more important to further analyze therole of serum response factor(SRF)and understand the molecular mechanism of diapause embryo restart.The significance of the study also provides reference materials for the research on the "molecular mechanism of the restart of diapause embryos of crustaceans and other invertebrates".