Cloning,Tissue Distribution and Temporal Expression Analysis of Synaptosome-associated Protein 29 Gene(SNAP29)in Cynoglossus semilaevis
The aim of this study was to analyze synaptosome associated protein 29 gene(SNAP29)in Cynoglos-sus semilaevis.The molecular biological characteristics,tissue distribution characteristics of SNAP29 gene and the temporal expression pattern after infection with Shewanella algae.The SNAP29 gene of C.semilaevis was amplified by RT-PCR,bioinformatics website and qRT-PCR,and its bioinformatics analysis and expression characteristics were studied.The results showed that the CDS region of SNAP29 gene was 789 bp in length,encoding 262 amino acids.The theoretical isoelectric point(pI)was 5.39,and the molecular weight was29.71781 ku.There were two typical SNARE domains.The secondary structure was mainly composed of α-helix and random coil,and the secondary structure and tertiary structure were basically the same.The results of multiple sequence alignment showed that the SNAP29 gene of C.semilaevis had the highest similarity with that of European perch.The SNAP29 gene was ex-pressed in all tissues of healthy C.semilaevis,with the highest expression in gill tissue and the lowest expression in brain tissue.After artificial infection of C.semilaevis with S.algae,the expression of SNAP29 gene was mainly up-regulated in intestinal tissue,heart tissue and brain tissue,and down-regulated in liver,head kidney and spleen tis-sue.SNAP29 gene was expressed in heart,brain,liver,intestinal tract,spleen,gill,middle kidney and head kidney of healthy C.semilaevis.The SNAP29 gene was differentially expressed in the intestine,brain,heart,liver,head kidney and spleen of C.semilaevis infected with S.algae.The above results indicate that SNAP29 may be involved in the body's immune response process or pathophysiological process.