The Fuction Of API5 In Dispause Embryo Restart Of Artemia Sinica

Apoptosis inhibitor 5,a gene with a remarkable ability of anti-apoptosis.Base on the provious research result of As-api5,this study make a futher analysis of the full cDNA sequence of As-api5 by bioinformatics method,which found that the putative As-API5 protein has a p I of 9.24 and has no signal peptide which suggests it's not a secretory protein.The hydrophobicity analysis of As-API5 indicated that the protein is hydrophilic.Result of PSORTII prediction showed that As-API5 contains twenty-seven phosphorylation sites,including twenty serine,four threonine,and three tyrosine sites.ISH showed As-api5 signals were observed in different locations during development periods of A.sinica,which indicated that the gene expressed widely in A.sinica and without specific expression in tissues and organs.Down regulation of As-api5 expression using a short interfering RNA resulted in increased mortality and embryo malformation of A.sinica.As-API5 was present in the pellet fraction which was founded by prokaryotic expression method.Result of western blot showed that As-API5,As-E2F1,As-Cyclin E,As-APAF1,As-Caspase9,As-CyclinA and As-CDK2 have the similar expression trend during different development periods of A.sinica.After the embryonic diapauses termination,embryo restarts,the differentiation and proliferation of cells become rapidly,the expression level was up regulated(0-10h),after that(15h-3d),the vital signs of A.sinica gradually become stable,the expression of As-API5 showed a down trend,and keep at a basic level.Under the stress of high salinity and low temperature condition,the expression of As-API5 showed a ascending trend.And related protein also presented a similar trend of As-API5 in western blot.The function of API5 in embryo restart process of A.sinica was analyzed,experimental results showed API5 played an crucial role in embryo restart and early embryonic development of A.sinica,especially in the regulation of cell cycle and apoptosis.And keep the balance of profliferation and apoptosis,in order to resist the unsuitable condition.