Share This Article:

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

Abstract Full-Text HTML XML Download Download as PDF (Size:1431KB) PP. 123-127
DOI: 10.4236/abb.2011.23019    4,101 Downloads   9,416 Views   Citations


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.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Chang, C. , Liu, X. and Chen, K. (2011) Molecular cloning, expression and characterization of a novel geneβ-N-acetylglucosaminidase from Bombyxmori. Advances in Bioscience and Biotechnology, 2, 123-127. doi: 10.4236/abb.2011.23019.


[1] Chen, H.Q., Chen, K.P., Yao, Q., Guo, Z.J. and Wang, L.L. (2007) Characterization of a late gene, ORF67 from bombyx mori nucleopolyhedrovirus. FEBS Letters, 581, 5836-5842. doi:10.1016/j.febslet.2007.11.059
[2] Nakazawa, H., Tsuneishi, E., Ponnuvel, K.M., Furukawa, S., Asaoka, A., et al. (2004) Antiviral activity of a serine protease from the digestive juice of Bombyx mori larvae against nucleopolyhedrovirus. Virology, 321, 154-162. doi:10.1016/j.virol.2003.12.011
[3] Ponnuvel, K.M., Nakazawa, H., Furukawa, S., Asaoka, A., Ishibashi, J., et al. (2003) A lipase isolated from the silkworm bombyx mori shows antiviral activity against nucleopolyhedrovirus. Journal of Virology, 77, 10725- 10729. doi:10.1128/JVI.77.19.10725-10729.2003
[4] Aumiller, J.J., Hollister, J.R. and Jarvis, D.L. (2006) Molecular cloning and functional characterization of beta-N-acetylglucosaminidase genes from Sf9 cells. Protein Expression and Purification, 47, 571-590. doi:10.1016/j.pep.2005.11.026
[5] Cattaneo, F., Pasini, M.E., Intra, J., Matsumoto, M., Briani, F., et al. (2006) Identification and expression analysis of Drosophila melanogaster genes encoding β-hexosaminidases of the sperm plasma membrane. Glycobiology, 16, 786-800. doi:10.1093/glycob/cwl007
[6] Nagamatsu, Y., Yanagisawa, I., Kimoto, M., Okamoto, E. and Koga, D. (1995) Purification of a chitooligosaccharidolytic beta-N-acetylglucosaminidase from Bombyx mori larvae during metamorphosis and the nucleotide sequence of its cDNA. Bioscience, Biotechnology, and Biochemistry, 59, 219-225. doi:10.1271/bbb.59.219
[7] Takahiro, O., Seiji, I., Hideki, S., Akihiro, U., Toshiki, T., et al. (2007) Molecular cloning and expression of two novel β-n-acetylglucosaminidases from silkworm bombyx mori. Bioscience, Biotechnology, and Biochemistry, 71, 1626-1635. doi:10.1271/bbb.60705
[8] Jarvis, D.L., Wills, L., Burow, G. and Bohlmeyer, D.A. (1998) Mutational analysis of the N-linked glycans on autographa californica nucleopolyhedrovirus gp64. Journal of Virology, 72, 9459-9469.
[9] Liu, X.Y., et al. (2010) Proteomic analysis of nucleopolyhedrovirus infection resistance in the silkworm, Bombyx mori (Lepidoptera: Bombycidae). Journal of Invertebrate Pathology, 105, 84-90.
[10] Li, X.H., Wu, X.F., Yue, W.F., Liu, J.M., Li, G.L., et al. (2006) Proteomic analysis of the silkworm (Bombyx mori L.) hemolymph during developmental stage. Journal of Proteome Research, 5, 2809-2814. doi:10.1021/pr0603093
[11] Kothari, H., Kumar, P. and Singh, N. (2006) Prokaryotic expression, purification, and polyclonal antibody production against a novel drug resistance gene of Leishmania donovani clinical isolate. Protein Expression and Purification, 45, 15-21. doi:10.1016/j.pep.2005.10.002
[12] Bade, M.L. and Wyatt, G.R. (1962) Metabolic conversions during putation of the cecropia silkworm. 1. Deposition and utilization of nutrient. Biochemical Journal, 83, 470-478.
[13] Zen, K.C., Choi, H.K., Krishnamachary, N., Muthukrishnan, S. and Kramer, K.J. (1996) Cloning, expression and hormonal regulation of an insect β-N-acetylglucosa- minidase gene. Insect Biochemistry and Molecular Biology, 26, 435-444. doi:10.1016/0965-1748(95)00111-5

comments powered by Disqus

Copyright © 2019 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.