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Advanced Protein Expression Using Bombyx mori Nucleopolyhedrovirus (BmNPV) Bacmid in Silkworm

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Short Views on Insect Genomics and Proteomics

Part of the book series: Entomology in Focus ((ENFO,volume 4))

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Abstract

An expression system using a Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid has been developed and used for efficient expression of protein using silkworm. Silkworm can sustain large-scale production of recombinant proteins due to its ease of rearing and scaling-up. Our chapter focuses on the modification of a BmNPV bacmid for a more efficient protein expression system. For example, we discuss how to achieve construction of a stronger promoter, less proteolytic degradation of expressed proteins, and a chaperone-coexpressed expression system. We describe the application of functional BmNPV particles purified from silkworm hemolymph to vaccines, antibody production, and transmembrane protein analysis. For human use, the major problem of proteins produced in silkworm is contamination by adventitious agents and protein quality. Of special concern is that N-glycosylation in silkworms is of a high-mannose type in most cases, which is different from the complex type found in mammals. We end by looking to future prospects for integration/applications of protein expression systems with silkworm biotechnology.

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Abbreviations

α4GnT:

Human α1,4-N-acetylglucosaminyltransferase

AcMNPV:

Autographa californica multiple nucleopolyhedrovirus

ADCC:

Antibody-dependent cell-mediated cytotoxicity

β3GnT:

Human β1,3-N-acetylglucosaminyltransferase

BiP:

Human heavy chain-binding protein

Bm:

Bombyx mori

BmNPV:

Bombyx mori nucleopolyhedrovirus

BmNPV CP− (CP−):

Cysteine protease-deficient BmNPV

BmNPV CP− Chi− (CP−Chi−):

Cysteine protease- and chitinase-deficient BmNPV

CNX:

Human calnexin

CRT:

Human calreticulin

CSFV:

Classical swine fever virus

DDS:

Drug delivery system

ELISA:

Enzyme-linked immunosorbent assay

ERp:

Human ERp57

FMDV:

Foot-and-mouth disease virus

GFP:

Green fluorescent protein

HA:

Hemagglutinin

Hsp70:

Heat shock protein 70

IgG:

Immunoglobulin G

IL-5:

Interleukin-5

LUV:

Large unilamellar vesicle

MSG:

Middle silk gland

PIT, 7d:

Post-injection time, 7 days

PSG:

Posterior silk gland

RSV:

Rous sarcoma virus

RV:

Rabies virus

SAG1:

Surface antigen 1

SRB:

Sulforhodamine B

SRS2:

Surface antigen 1-related sequence 2

ST6Gal1:

α2,6-sialyltransferase 1

VLP:

Virus-like particle

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Authors and Affiliations

  1. Green Chemistry Research Division, Research Institute of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan

    Tatsuya Kato & Enoch Y. Park

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  1. Tatsuya Kato
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  2. Enoch Y. Park
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Correspondence to Enoch Y. Park .

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Editors and Affiliations

  1. Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, Kansas, USA

    Chandrasekar Raman

  2. Biological Sciences Department Center for Biotech. and Life Sciences, University of Rhode Island, Kingston, Rhode Island, USA

    Marian R. Goldsmith

  3. Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut, USA

    Tolulope A. Agunbiade

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Kato, T., Park, E.Y. (2016). Advanced Protein Expression Using Bombyx mori Nucleopolyhedrovirus (BmNPV) Bacmid in Silkworm. In: Raman, C., Goldsmith, M., Agunbiade, T. (eds) Short Views on Insect Genomics and Proteomics. Entomology in Focus, vol 4. Springer, Cham. https://doi.org/10.1007/978-3-319-24244-6_7

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