Cloning And Function Study Of Flotillins From Dugesia Japonica

Flotillins are markers associated with lipid raft protein, and they are widely distributed in animals,also they are very conserved protein. During the development of the individual, Flotillins are extensivelyinvolved in many physiological functions, such as neurite growth, endocytosis, tumor formation, theformation of germ cells, insulin regulation.In this article, we took Dugesia japonica as experimental material, used RACE to get twofull-length cDNA sequence of Djflotillins. The full-length cDNA sequence of the Djflotillin-1is1336bp,its ORF is1260bp, codes420amino acids. The full-length cDNA sequence of the Djflotillin-2is1588bp,its ORF is1296bp, codes430amino acids. There is a SPFH domain between37and171amino acid ofDjFlotillin-1. Also there is a SPFH domain between40and176amino acid of DjFlotillin-2.Used bioinformatics techniques to analyze the physicochemical properties of these two proteins.The molecular weight of DjFlotillin-1is46.7Kda, Isoelectric point is6.94. The molecular weight ofDjFlotillin-2is46.9Kda, Isoelectric point is5.19. Both of them are hydrophilic protein, have severalphosphorylation sites, don’t have signal peptide and transmembrane region.By Real-time PCR to study the expression trend of the two genes in regeneration. The resultsshowed that: they had the same expression trend, their mRNA had elevated expression levels of the firstday, then lowered, since the second day to the fifth day, their expression trend were increased and reachedthe highest value in the fifth day, then lowered again in the sixth day and seventh day. So we speculate thatthese two proteins may promote the differentiation of neural stem cells in the early regeneration of the head,and also paly an important role in the formation and function recovery of axons in the middle and latestage.