scholarly journals Alkaline Protease Associated with Virus Particles of a Nuclear Polyhedrosis Virus: Assay, Purification, and Properties

1978 ◽  
Vol 26 (1) ◽  
pp. 84-92 ◽  
Author(s):  
C. C. Payne ◽  
J. Kalmakoff
Keyword(s):  
Alkaline Protease ◽  
Nuclear Polyhedrosis Virus ◽  
Virus Particles ◽  
Virus Assay ◽  
Purification And Properties ◽  
Polyhedrosis Virus ◽  
Nuclear Polyhedrosis

A VIRUS DISEASE OF ECTROPIS CREPUSCULARIA SCHIFF. (GEOMETRIDAE: LEPIDOPTERA)

1967 ◽  
Vol 13 (7) ◽  
pp. 855-858 ◽  
Author(s):  
Oswald N. Morris
Keyword(s):  
Dna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Nuclear Polyhedrosis Virus ◽  
Virus Disease ◽  
Disease Process ◽  
Virus Diseases ◽  
Virus Particles ◽  
Polyhedrosis Virus ◽  
Nuclear Polyhedrosis

The morphology of a nuclear polyhedrosis virus of Ectropis crepuscularia Schiff., the histopathology of the disease, and deoxyribonucleic acid (DNA) synthesis during the disease process were studied. The polyhedral inclusions measure 1.4 μ in diameter with a range of 0.8–2.1 μ. The virus particles are rods measuring 319 mμ by 79 mμ with a range of 280–360 mμ by 70–110 mμ. DNA labelling and nuclear swelling follow the course of previously described polyhedrosis virus diseases.

Purification and properties of a nuclear polyhedrosis virus from Choristoneura fumiferana

1975 ◽  
Vol 21 (8) ◽  
pp. 1224-1231 ◽  
Author(s):  
Basil M. Arif ◽  
Keith W. Brown
Keyword(s):  
Nuclear Polyhedrosis Virus ◽  
Alkali Treatment ◽  
Choristoneura Fumiferana ◽  
Spruce Budworm ◽  
Potassium Tartrate ◽  
Purification And Properties ◽  
Nonionic Detergent ◽  
Polyhedrosis Virus ◽  
Nuclear Polyhedrosis ◽  
Equilibrium Centrifugation

The polyhedral inclusion bodies of a nuclear polyhedrosis virus from the spruce budworm Choristoneura fumiferana were purified by fluorocarbon treatment, sucrose columns, sucrose density gradients, and equilibrium centrifugation. The preparation yields a single and homogeneous band in potassium tartrate gradients at a density of 1.193 g/ml. The virions were released from the polyhedra by alkali treatment and purified by either rate zonal or equilibrium centrifugation in sucrose gradients. The purified virions, which band at a density of 1.265–1.300 g/ml in sucrose, were infectious to the spruce budworm larvae and their mean LD50 was 1.09 ± 0.1 μg of virus. However, viral nucleocapsids, released by a nonionic detergent, Nonidet P-40, had a density of 1.315 g/ml and were noninfectious by either per os inoculation or intrahemocoelic injection.

scholarly journals Bombyx mori Pupae Efficiently Produce Recombinant AAV2/HBoV1 Vectors with a Bombyx mori Nuclear Polyhedrosis Virus Expression System

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 704
Author(s):  
Qian Yu ◽  
Pengfei Chang ◽  
Xiaoxuan Liu ◽  
Peng Lü ◽  
Qi Tang ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Bombyx Mori ◽  
Nuclear Polyhedrosis Virus ◽  
Expression System ◽  
Efficient Production ◽  
Human Bocavirus ◽  
Adeno Associated Virus ◽  
Silkworm Pupae ◽  
Polyhedrosis Virus ◽  
Nuclear Polyhedrosis

Recombinant adeno-associated virus (AAV) vectors have broad application prospects in the field of gene therapy. The establishment of low-cost and large-scale manufacturing is now the general agenda for industry. The baculovirus-insect cell/larva expression system has great potential for these applications due to its scalability and predictable biosafety. To establish a more efficient production system, Bombyx mori pupae were used as a new platform and infected with recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV). The production of a chimeric recombinant adeno-associated virus (rAAV) serotype 2/human bocavirus type-1 (HBoV1) vector was used to evaluate the efficiency of this new baculovirus expression vector (BEV)–insect expression system. For this purpose, we constructed two recombinant BmNPVs, which were named rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP. The yields of rAAV2/HBoV1 derived from the rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP co-infected BmN cells exceeded 2 × 104 vector genomes (VG) per cell. The rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP can express stably for at least five passages. Significantly, rAAV2/HBoV1 could be efficiently generated from BmNPV-infected silkworm larvae and pupae at average yields of 2.52 × 1012 VG/larva and 4.6 × 1012 VG/pupa, respectively. However, the vectors produced from the larvae and pupae had a high percentage of empty particles, which suggests that further optimization is required for this platform in the future. Our work shows that silkworm pupae, as an efficient bioreactor, have great potential for application in the production of gene therapy vectors.

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