Genes Cloning And Expression Analysis Of β GRP、Immulectins From The Chinese Oak Silkmoth,Antheraea Pernyi

Insect immunity is different from vertebrate, they have no T cell.B cell and complement system. Insects resist invasion of foreign pathogenic microorganisms, in addition to rely on itself special structures like the exoskeletons. peritrophic membrane, mainly rely on humoral immune and cellular immune to destroy invading organisms. Among them, pattern recognition receptor mediated by the humoral immune defense system plays an important role in insects.In this paper, βGRP, immunlectin-5and immunlectin-A genes of antheraea pernyi were cloned, and expression profile which induced by different microoganism were investigated. The results showed that open reading frame of PGRP is consisted of490amino acids, with a predicted signal peptide of17amino acids, it contains a conservative the structural domain with glycosidic Hydrolase activity（Glyco_hydro₁6）. Immunlectin-5has an open reading frame（ORF） of522bp and codes for174amino acids.with a predicted signal peptide of29amino acids and immunlectin-A has an open reading frame（ORF）of951bp and codes for317amino acids.with a predicted signal peptide of24amino acids. These two genes contain conservative carbohydrate recognition structure domain （CRD）, can identify specific polysaccharide and participate in insect immunity. After antheraea pernyi pupa were infected with Bacillus subtilis, Escherichia coli ToplO. antheraea pernyi nuclear polyhedrosis virus （ApNPV） and Saccharomyces. cerevisiae W303-1A respectively, the results showed that injected different pathogenic microorganisms, these genes have different expression level in different tissuses（midgut. fatbody, epidermis, testicle, hemocytes）. The high level expression of Apβ3GRP can be induced by Bacillus subtilis, it has the highest expression level in fatbody and midgut.Immunlectin-5can be induced by Bacillus subtilis. Escherichia coli Top1O and S. cerevisiae W303-1A, the expression level of epidermis is higher than other tissuses. The expression of immunlectin-A is upregulated after injected by Bacillus subtilis. Escherichia coli Top1O. Antheraea pernyi nuclear polyhedrosis virus （ApNPV） and S. cerevisiae W303-1A. especially in the fatbody it has the highest expression. These data indicate that ApβGRP can response gram-positive bacterium through recognizing molecular patterns associated with phosphorus acid of gram-positive organisms, it is play an important role in Antheraea pernyi immune system. Immulectin-5and Immunlectin-A are widely antibacterial activity, they also play an important role in Antheraea pernyi immune system.In addition, we use inhibitors（NF/kB inhibitor and JAK inhibitor） to research the mechanism of three receptors, the results suggest:Ap(3GRP. immulectin-5and immulectin-A regulate the expression of AMPs through NF/kB factor. However, we need to further study about their antiviral mechanism.