动物造模 z-bio 2022-03-07 310

　　Objective To obtain the full-length coding sequence of junctional adhesion molecule A in tree ferrets and analyze its molecular characteristics, and to explore the mechanism of JAM-A as a receptor for reovirus invading primary lung cells of tree ferrets.

　　Methods Total RNA was extracted from healthy ferret tissues, and JAM-4 gene was obtained by reverse transcription PCR and cDNA end rapid cloning technology; the encoded amino acids were analyzed by Unipro UGENE, and a phylogenetic tree was constructed by using MEGA6.0 software maximum likelihood method; real-time fluorescence quantitative PCR The expression and distribution of JAM-A in 25 tissues and blood were detected; the primary lung epithelial cells were subjected to receptor-specific antibody blocking treatment, and then immunofluorescence was used to observe the effect of viral antigen on reosomes. Effects of viral infection.

　　Results The full-length cDNA sequence of JAM-A gene was obtained, 2962bp; phylogenetic tree evolution analysis showed that JAM-4 gene was more closely related to humans than rodents; JAM-A molecule was widely expressed in the peripheral tissues of arborvitae, while in Higher levels of expression in the respiratory and digestive tracts. Specific antibodies acting on cells significantly reduced the immunofluorescence integral absorbance of viral antigens (P<0.0001).

　　Conclusion The gene sequence of JAM-A was cloned and analyzed for the first time, and it was verified that JAM-A molecule is the main receptor of reovirus invasion tree, suggesting that tree can be used as a new animal model of reovirus.