Abstract
Using human genomic DNA as a template, the human insulin gene was cloned and used to construct various reBmMNPVbacmids. Cysteine protease gene deletion (CPD-BmMNPV bacmid) and cysteine protease- and chitinase-deficient (CPPD- BmMNPV bacmid) baculoviruses were used to express both native and FLAG-tagged human insulin. Silkworm larvae were infected with the above recombinant bacmid DNAs, and the expressed insulin was purified and identified from infected silkworm haemolymph. The highest expression was shown with the CPPD- BmMNPV bacmid, which was about two times that of the wild type of reBmMNPVbacmid, reaching 15.827 ng/ml haemolymph.
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Acknowledgements
The works were supported by the National Basic Research Program of China under grand No. 2005CB121003 and the National Natural Science Foundation of China (NSFC: 30972141/C120110) and the Hi-Tech Research and Development Program of China (No. 2008AA10Z132 and No. 2006AA10A119). The work was conducted in the Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Japan.
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Yue, Wf., Zhou, F., Hu, Jb. et al. Human insulin gene expressing with Bombyx mori multiple nucleopolyhedrovirus (BmMNPV) expression system. World J Microbiol Biotechnol 27, 393–399 (2011). https://doi.org/10.1007/s11274-010-0470-z
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DOI: https://doi.org/10.1007/s11274-010-0470-z