Cloning, Expression And Bioinformatics Analysis Of CD81Gene From Lampetra Japonica

CD81is a member of the tetraspanin family that has been described to have a key role incell signal transduction,cell migration of tumor and immune cells and so on.CD81is acomponent of B cell and T cell receptor, the interaction between B cell and T cell can inhibitthe transmission of information.CD81combined with CD4、CD8、CD19、CD21、HLA-DR、α3β1, and regulation hepatitis C virus(HCV) infection host cell.The present studies mainlyconcentrated in higher vertebrates.But the biological function of CD81in primitivevertebrates is still not clear.In this paper,by searching and bioinformatics analysis the sequences of cDNA,get a highhomology with the CD81sequence,which can be translated CD81protein open readingframe.According to the sequence information and enzyme sites designed primers, then takethe total RNA as a template, using PCR amplification method get the fragment of CD81, andsuccessfully build pET42a (+)-CD81prokaryotic expression vector.The alignment analysisof CD81amino acid sequence and construct phylogenetic tree.In addition, the analysis ofsecondary structure domain,tertiary structure domain prediction,hen water prediction,disulfidebond prediction,antigen epitope and conservative motif prediction.Through performing semi-quantitative PCR assay, we analyzed the relative expressionlevel of CD81in heart, fat body, kidney, liver, gill and leukocyte. Analysis the expressionlevel changes of CD81in control group and stimulating group.In conclusion, Lamprey between vertebrates and invertebrates.The discovery of CD81contribute to explore CD81regulating lampreys hematopoietic system and the evolution ofCD81.