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

Molecular cloning, expression and characterization of a novel geneβ-N-acetylglucosaminidase from Bombyxmori
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摘要 Previously, we have reported that a gene encoding Bombyxmoriβ-N-acetylglucosaminidase 2 (BmGlcNAcase2) 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 BC9 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 BmGlcNAcase2 was mainly located to the cytoplasm or some structure in cytoplasm. Previously, we have reported that a gene encoding Bombyxmoriβ-N-acetylglucosaminidase 2 (BmGlcNAcase2) 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 BC9 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 BmGlcNAcase2 was mainly located to the cytoplasm or some structure in cytoplasm.
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出处 《Advances in Bioscience and Biotechnology》 2011年第3期123-127,共5页 生命科学与技术进展(英文)
关键词 Bombyxmori β-N-acetylglucosaminidase2 WESTERN BOLT Sub-Cellular Localization Bombyxmori β-N-acetylglucosaminidase2 Western Bolt Sub-Cellular Localization
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