Molecular cloning, expression and characterization of a novel geneβ-N-acetylglucosaminidase from Bombyxmori

Abstract

Previously, we have reported that a gene encoding Bombyxmoriβ-N-acetylglucosaminidase 2 (BmGlcNA case2) has been identified differentially expressed in the midgut of Bombyxmori strain NB resistant to nucleopolyhedrovirus (BmNPV), strain 306 susceptible to NPV and a near isogenic line BC 9 with similar genetic background to 306 but resistant to NPV by two-dimensional gel electrophoresis (2-DE). To get more knowledge about the relationship between β-N--acetylglucosaminidase and the resistance of NPV, in this study, the 1542 bp open reading frame of a putative bombyxmoriβ-N-acetylglucosaminidase 2 gene (BmGlcNAcase2) was amplified from a pool of bombyxmoricDNAs and inserted into the prokaryotic expression plasmid pET-30a(+). Western blotting analysis showed that BmGlcNAcase2 was expressed in hemolymph, ovary, testis, fat body, trachea, midgut and silk gland of fifth instar larvae respectively. Immunofluoresence analysis indicated that BmGlcNA case2 was mainly located to the cytoplasm or some structure in cytoplasm.

The Differential Expression of BmGlcNAcase2 in Strains of Bombyx mori (Lepidoptera: Bombycidae) With Different Susceptibility to Bombyx mori (Lepidoptera: Bombycidae) Nucleopolyhedrovirus Infection

“…Today, strains NB and 306 (the strain which is susceptible to BmNPV) are often used in comparative research of BmNPV resistance. In our previous two-dimensional gel electrophoresis study, our group determined that the expression of β-N-acetylglucosaminidase 2 (GlcNAcase2) is different between strains NB and 306 ( Chang 2011 ). Here, we investigated whether the difference in resistance to BmNPV is related to GlcNAcase 2.…”