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- Title
The use of defective Bombyx mori nucleopolyhedrovirus genomes maintained in Escherichia coli for the rapid generation of occlusion-positive and occlusion-negative expression vectors.
- Authors
Yeon Ho Je; Jin Hee Chang; Mi Hyang Kim; Jong Yul Roh; Byung Rae Jin; O'Reilly, David R.
- Abstract
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.
- Subjects
GENETIC vectors; SILKWORMS; ESCHERICHIA coli; RECOMBINANT viruses; GENOMICS; GENES
- Publication
Biotechnology Letters, 2001, Vol 23, Issue 21, p1809
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1023/A:1012475004212