Baculovirus-Mediated Production of the Human Growth Hormone in Larvae of the Silkworm, Bombyx mori

Kadono-Okuda, Keiko; Yamamoto, Masanori; Higashino, Yusuke; Taniai, Kiyoko; Kato, Yosuke; Chowdhury, Subrata; Xu, Jinhua; Choi, Su Kyung; Sugiyama, M.; Nakashima, Kunio; Maeda, Susumu; Yamakawa, Minoru

doi:10.1006/bbrc.1995.2144

Cited by 32 publications

(20 citation statements)

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“…It is difficult to speculate about the reasons underlying the higher sensitivity of VP3 to proteolysis when expressed in insect cells. Degradation of proteins expressed in insect cells with rBVs has been reported frequently (17,34,39). VP3 C-terminal trimming might be caused by a protease of either cellular or viral origin.…”

Section: Discussionmentioning

confidence: 99%

The Oligomerization Domain of VP3, the Scaffolding Protein of Infectious Bursal Disease Virus, Plays a Critical Role in Capsid Assembly

Maraver¹,

Oña²,

Abaitua³

et al. 2003

J Virol

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Infectious bursal disease virus (IBDV) capsids are formed by a single protein layer containing three polypeptides, pVP2, VP2, and VP3. Here, we show that the VP3 protein synthesized in insect cells, either after expression of the complete polyprotein or from a VP3 gene construct, is proteolytically degraded, leading to the accumulation of product lacking the 13 C-terminal residues. This finding led to identification of the VP3 oligomerization domain within a 24-amino-acid stretch near the C-terminal end of the polypeptide, partially overlapping the VP1 binding domain. Inactivation of the VP3 oligomerization domain, by either proteolysis or deletion of the polyprotein gene, abolishes viruslike particle formation. Formation of VP3-VP1 complexes in cells infected with a dual recombinant baculovirus simultaneously expressing the polyprotein and VP1 prevented VP3 proteolysis and led to efficient virus-like particle formation in insect cells.

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Section: Discussionmentioning

confidence: 99%

The Oligomerization Domain of VP3, the Scaffolding Protein of Infectious Bursal Disease Virus, Plays a Critical Role in Capsid Assembly

Maraver¹,

Oña²,

Abaitua³

et al. 2003

J Virol

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“…Expression of hGH has also been reported in diverse organisms, such as yeast (Apte-Deshpande et al 2009), Bacillus subtilis (Ozdamar et al 2009), rabbits (Lipiński et al 2003), baculovirus (Cholin et al 1996), silkworms (Kadono-Okuda et al 1995;Sumathy et al 1996), pigs (Hens et al 2000), sheep (Sanchez et al 2004), CHO animal cells (Kunaparaju et al 2005), cows (Salamone et al 2006) and mice (von Waldthausen et al 2008). The possibility of producing recombinant proteins in plant systems, the least expensive biomass, is another option being considered as a means to lower the costs associated with the large-scale production of pharmaceuticals and industrial molecules (Kusnadi et al 1997;Daniell et al 2001;Ma et al 2003;Twyman et al 2003).…”

Section: Introductionmentioning

confidence: 94%

Expression of functional recombinant human growth hormone in transgenic soybean seeds

et al. 2010

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We produced human growth hormone (hGH), a protein that stimulates growth and cell reproduction, in genetically engineered soybean [Glycine max (L.) Merrill] seeds. Utilising the alpha prime (α') subunit of β-conglycinin tissue-specific promoter from soybean and the α-Coixin signal peptide from Coix lacryma-jobi, we obtained transgenic soybean lines that expressed the mature form of hGH in their seeds. Expression levels of bioactive hGH up to 2.9% of the total soluble seed protein content (corresponding to approximately 9 g kg(-1)) were measured in mature dry soybean seeds. The results of ultrastructural immunocytochemistry assays indicated that the recombinant hGH in seed cotyledonary cells was efficiently directed to protein storage vacuoles. Specific bioassays demonstrated that the hGH expressed in the soybean seeds was fully active. The recombinant hGH protein sequence was confirmed by mass spectrometry characterisation. These results demonstrate that the utilisation of tissue-specific regulatory sequences is an attractive and viable option for achieving high-yield production of recombinant proteins in stable transgenic soybean seeds.

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“…Among a number of promising malaria vaccines, Plasmodium falciparum merozoite surface protein 1 (MSP-1) is a leading candidate for a human erythrocytic malaria vaccine. MSP-1 is synthesized during schizogony as a 195-kDa glycoprotein (19) and is proteolytically processed into fragments of 83, 30, 38, and 42 kDa, designated MSP-1 83 , MSP-1 30 , MSP-1 38 , and MSP-1 42 , respectively (20,39). During erythrocytic invasion, MSP-1 42 is cleaved to yield 33-and 19-kDa fragments (MSP-1 33 and MSP-1 19 , respectively) (4,5).…”

mentioning

confidence: 99%

“…To deal with these shortcomings, we chose to express MSP-1 42 in silkworm larvae by using the silkworm-specific baculovirus Bombyx mori nuclear polyhedrosis virus (BmNPV). By using this in vivo expression system, a number of recombinant proteins of pharmaceutical and agricultural importance, including human interferons (13,40), human growth hormone (30,52), human macrophage colony-stimulating factor (46), human granulocyte-macrophage colony-stimulating factor (47), viral proteins (54,58), and grass carp growth hormone (18), have been successfully expressed with biological activities comparable to those of the native counterparts. The expression levels of these recombinant proteins vary, but up to 13 mg/larva has been reported (43).…”

mentioning

confidence: 99%

In Vivo Expression and Immunological Studies of the 42-Kilodalton Carboxyl-Terminal Processing Fragment ofPlasmodium falciparumMerozoite Surface Protein 1 in the Baculovirus-Silkworm System

et al. 2002

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The 42-kDa carboxyl-terminal processing fragment of Plasmodium falciparum merozoite surface protein 1 (MSP-1 42 ) is an anti-erythrocytic stage malaria vaccine candidate. In this study, MSP-1 42 was expressed by using the Bombyx mori nuclear polyhedrosis virus-silkworm expression system, and the antigenicity and immmunogenicity of the recombinant protein, Bmp42, were evaluated. The average yield of Bmp42, as determined by a sandwich enzyme-linked immunosorbent assay (ELISA), was 379 g/ml of infected silkworm hemolymph, which was >100-fold higher than the level attainable in cell culture medium. N-terminal amino acid sequencing revealed that Bmp42 was correctly processed in silkworm cells. Data from immunoblotting, as well as from the inhibition ELISA, suggested that the conformational B-cell epitopes of MSP-1 42 were recreated in Bmp42. Immunization of rabbits with Bmp42 in complete Freund's adjuvant generated high-titer antibody responses against the immunogen. Specificity analyses of the anti-Bmp42 antibodies using several recombinant MSP-1 19 proteins expressing variant and conserved B-cell epitopes suggested that the anti-Bmp42 antibodies recognized primarily conserved epitopes on MSP-1 19 . Furthermore, the anti-Bmp42 antibodies were highly effective in inhibiting the in vitro growth of parasites carrying homologous or heterologous MSP-1 42 . Our results demonstrated that the baculovirus-silkworm expression system could be employed to express biologically and immunologically active recombinant MSP-1 42 at elevated levels; thus, it is an attractive alternative for producing a protective MSP-1 42 vaccine for human use.

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Baculovirus-Mediated Production of the Human Growth Hormone in Larvae of the Silkworm, Bombyx mori

Cited by 32 publications

References 0 publications

The Oligomerization Domain of VP3, the Scaffolding Protein of Infectious Bursal Disease Virus, Plays a Critical Role in Capsid Assembly

The Oligomerization Domain of VP3, the Scaffolding Protein of Infectious Bursal Disease Virus, Plays a Critical Role in Capsid Assembly

Expression of functional recombinant human growth hormone in transgenic soybean seeds

In Vivo Expression and Immunological Studies of the 42-Kilodalton Carboxyl-Terminal Processing Fragment ofPlasmodium falciparumMerozoite Surface Protein 1 in the Baculovirus-Silkworm System

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