The use of defective Bombyx mori nucleopolyhedrovirus genomes maintained in Escherichia coli for the rapid generation of occlusion-positive and occlusion-negative expression vectors

A system is described for the rapid generation of Bombyx mori nucleopolyhedrovirus (BmNPV)-based expression vectors. A series of novel BmNPV genomes, that include a mini-F replicon and therefore can be maintained in Escherichia coli, have been generated. These genomes lack a portion of the essential...

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Bibliographic Details
Published in:Biotechnology letters 2001-01, Vol.23 (21), p.1809-1817
Main Authors: Je, Yeon Ho, Chang, Jin Hee, Hyang Kim, Mi, Yul Roh, Jong, Rae Jin, Byung, O'reilly, David R
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
E coli
Escherichia coli
Fundamental and applied biological sciences. Psychology
Genes
Genetic engineering
Genetic technics
Insects
Larvae
Methods. Procedures. Technologies
Microbiology
Nuclear polyhedrosis virus
Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Summary:A system is described for the rapid generation of Bombyx mori nucleopolyhedrovirus (BmNPV)-based expression vectors. A series of novel BmNPV genomes, that include a mini-F replicon and therefore can be maintained in Escherichia coli, have been generated. These genomes lack a portion of the essential ORF1629 gene and cannot replicate independently in insect cells. However, they can be used as parental genomes for the generation of expression vectors by cotransfection with a transfer plasmid that includes an intact ORF1629. Only recombinant viruses that have acquired the ORF1629 gene from the transfer vector, and have therefore also acquired the foreign gene of interest, can replicate after cotransfection. Parental genomes with and without a polyhedrin gene are described, enabling the generation of occlusion-positive and occlusion-negative recombinant viruses. Occlusion-positive expression vectors enable the oral infection of B. mori larvae and can therefore be used for the mass production of a foreign protein in infected insects.[PUBLICATION ABSTRACT]
ISSN:0141-5492
1573-6776
DOI:10.1023/A:1012475004212