Browse > Article

Rapid Expression of Bm46 in Bombyx mori Cell Lines, Larvae and Pupae  

Wang, Haiyan (Institute of Life Sciences, Jiangsu University)
Chen, Keping (Institute of Life Sciences, Jiangsu University)
Guo, Zhongjian (Institute of Life Sciences, Jiangsu University)
Yao, Qin (Institute of Life Sciences, Jiangsu University)
Wang, Qiang (Institute of Life Sciences, Jiangsu University)
Mu, Runhong (College of medical technology, Jiangsu University)
Publication Information
International Journal of Industrial Entomology and Biomaterials / v.15, no.1, 2007 , pp. 35-38 More about this Journal
Abstract
In this study, ORF 46 of Bombyx mod nucleopolyhedrovirus(Bm46) fused with EGFP was expressed in Bombyx mod cell lines, larvae and pupae by BmNPV Bacmid system. Bm46 and EGFP were cloned into donor plasmid pFastBacHTb, which was transformed to competent DH10B cells containing helper and BmNPV bacmid by site-specific transposition. Recombinant bacmid was used to transfected BmN-4 cells to produce the recombinant baculovirus vBm-Bm46-EGFP. Recombination virus was injected into silkworm larvae and pupae. The expression of the fusion protein was monitored by examining green fluorescence using a fluorescent microscope. Intense fluorescence in cells and silkworm was observed at 4 days post-infection, indicating the Bm46-EGFP fusion gene was expressed successfully.
Keywords
Bm46; expression; Baculovirus; BmNPV bacmid; Bombyx mori larvae; Pupae;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Khurad, A.M. Mahulikar, A. Rathod, M.K. Rai, M.M. Kanginakudru, S. and Nagaraju, J. (2004) Vertical transmission of nucleopolyhedrovirus in the silkworm, Bombyx mori L. J of Invertebr Pathol 87, 8-15   DOI   ScienceOn
2 Luckow, V.A. (1991) Cloning and expression of heterologous genes in insect cells with baculovirus vectors, in: Recombinant DNA Technology and Applications. Prokop, A. Bajpai, R.K. Ho, C.S. (eds.), pp. 97-152, McGraw-Hill, New York
3 Motohashi, T. Shimojima, T. Fukagawa, T. Maenaka, K. and Park, E.Y. (2005) Efficient large-scale protein production of larvae and pupae of silkworm nuclear polyhedrosis virus (BmNPV) bacmid system. Biochem. Biophys. Res. Commun. 326, 564-569   DOI   ScienceOn
4 O'Reilly D.R., Miller L.K., Luckow V.E. (1992) Baculovirus Expression Vectors: A Laboratory Manual. Freeman, New York
5 Enoch Y. Park, Ayano Kageshima, Mi-Sun Kwon, and Tatsuya Kato (2007) Enhanced production of secretory $\beta$ 1,3-Nacetylglucosaminyltransferase 2 fusion protein into hemolymph of Bombyx mori larvae using recombinant BmNPV bacmid integrated signal sequence. J Biotechnol. 129, 681-688   DOI   ScienceOn
6 Rucker, E. Schneider, G. Steinhauser, K. Lower, R. Hauber, J. and Stauber, R.H. (2001) Rapid evaluation and optimization of recombinant protein production using GFP tagging, Protein Exp. Purif. 21, 220-223   DOI   ScienceOn
7 Kenichi Moto, Hisao Kojima, Masaaki Kurihara, Masafumi Iwami, and Shogo Matsumoto (2003) Cell-specific expression of enhanced green fluorescence protein under the control of neuropeptide gene promoters in the brain of the silkworm, Bombyx mori, using Bombyx mori nucleopolyhedrovirus-derived vectors. Insect Biochem Molec 33, 7-12   DOI   ScienceOn
8 Gomi, S. Majima and K. Maeda S. (1999) Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. J. Gen. Virol, 80, 1323-1337   DOI
9 Luckow, V.A. Lee, S.C. Barry, G.F. Olins P.O. (1993) Efficient generation of infectious recombinant baculoviruses by sitespecific transposon mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli, J. Virol. 67, 4566-4579