Bioinformatics And Expression Analysis Of AsHPO And AsYki Genes Of Artemia Sinica

Artemia sinica is a small crustacean that generally lives in salt lake or seas and contains a variety of nutrients,making it a good food for aquaculture.The growth and development of A.sinica are affected by a variety of factors,and it does not have acquired immunity.In the face of the complex environment,it is overcome by innate immunity.It has been reported that the growth of organs and tissues,as well as the division and differentiation of cells are closely related to the Hippo signaling pathway.At present,research on the Hippo signaling pathway is mainly focused on the occurrence of human tumors and the model organisms Drosophila,Bombyx mori,etc.However,there has been little progress in the aquatic crustacean A.sinica.In this study,A.sinica was used as the experimental material,and the expression patterns of As HPO and As Yki genes and their relationship were explored at the molecular level by using Real-time fluorescent PCR,Western blot,Immunehistochemistry,and CO-Immunocoprecipitation.The main research and results are as follows:1.Expression of As HPO and As Yki genes m RNA in A.sinica at different stagesRT-q PCR was used to determine the m RNA expression levels of As HPO and As Yki at different development stages.The results showed that with the growth and development of A.sinica,the m RNA expression of As HPO and As Yki tended to decrease,and both were the highest in the egg stage.However,there was a difference between As HPO and As Yki at the same development stage,and the expression of As HPO was increased in the later nauplius stage,while that of As Yki was decreased.These results indicated that As HPO and As Yki were closely related to the growth and development of A.sinica,and the specific mechanism required further study.2.Effects of stress environment on As HPO and As Yki of A.sinicaBacterial stress,temperature stress and salinity stress were applied to A.sinica,and the m RNA expression of As HPO and As Yki stress environment was measured by RT-q PCR.Under bacterial stress,when Vibrio harveyi stimulated A.sinica,the expression levels of As HPO and As Yki increased with the increase of bacterial concentration.When Micrococcus lysodeikticus stimulated A.sinica,the expression of As HPO was inhibited and the inhibition effect was decreased with the increase of bacterial concentration,while the expression of As Yki was decreased with the increase of bacterial concentration and finally expression inhibition occurred.Under temperature stress,the expression of As HPO tended to increase with the increase of temperature,while the expression of As Yki showed the opposite trend.Under salinity stress,the changes of As HPO expression were not significant,but the expression of As Yki showed a downward trend with the increase of salinity.The change in the expression of As Yki was more significant than that of As HPO in the stress environment.3.Detection of protein interaction between As HPO and As Yki of A.sinicaThe interaction between As HPO and As Yki was detected by CO-Immunoprecipitation(CO-IP)method,and identified by Western-blot method.Results show that As HPO and As Yki did not interact.4.Localization of As HPO and As Yki protein in A.sinica during development stagesThe localization of As HPO and As Yki protein in the development of A.sinica was detected by Immunohistochemistry.The results showed that both As Yki and As HPO expressed in A.sinica at all stages,and mainly located in the head and chest.However,the distribution of As Yki protein was much wider than that of As HPO protein,indicating that As Yki protein had more critical biological functions.