Cloning And Preliminary Study On Function Of Caveolin-1 Gene In Artemia

Caveolins are the signature proteins of specialized invaginations of the plasma membrane named caveolae, which function to regulate signal transduction within the cell. Caveolin-1 is a principal structural component of caveolae. The amino acid sequence of Caveolin-1 scaffolding domain allows this protein to interact and inhibit or activate with signalling molecules such as epidermal growth factor receptor, endothelial nitric-oxide synthase, G-proteins, c-Src-like kinases, protein kinase C.Artemia sinica belongs to Phylum Arthropoda, Class Crustacea, Subclass Branchiopoda, Order Anostaca, Family Artemiidae and Genus Artemia rtemia. It is widely distributing in salterns and salt lakes and powerful adaptation to extreme environments. Artemia can giving birth to nauplii by the ovoviviparous pathway, but releases encysted embryos by the oviparous pathway in the circumstances and condition of environmental stresses. The deliquescing of embryonic diapause and development of Artemia encysted embryos are to study the hot point in Developmental Biology.In our study, we cloned the full length of caveolin-1 gene cDNA through RACE technique, and made bioinformatic analysis with the sequence. We detected the expression in different development stage and environmengtal stresses to research the caveolin-1 function in Artemia by Real-time PCR. The results as follow:(1)A full length Artemia caveolin-1 cDNA of 974bp was cloned by RACE, and the caveolin-1 cDNA revealed a 672bp open reading frame(ORF).(2)By the bioinformatic analysis, the ORF encodes a protein of 224 amino acids, which belongs to Caveolins superfamily. Predicted molecular weight is 25.4KDa, theoretical isoelectric point is 5.48.(3)Under different salt concentrations and stimulation by bacteria stress shows that the caveolin-1 gene is highly expressed in environment of 0 hour, high salty and bacteria by quantitative Real-time PCR.(4) To sum up the above arguments, Artemia caveolin-1 may have the potential function in innate immunity, embryonic developmental regulation and cell cycle regulation.