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Nucleic Acids Research, 2003, Vol. 31, No. 2 e6
© 2003 Oxford University Press

Improving baculovirus recombination

Yuguang Zhao, David A. G. Chapman and Ian M. Jones*

Animal and Microbial Sciences, University of Reading, Whiteknights, Reading RG6 6AJ, UK

*To whom correspondence should be addressed. Tel/Fax: +44 118 931 8902; Email: i.m.jones{at}reading.ac.uk
Present address:
Yuguang Zhao, Institute of Molecular Medicine, Oxford University, John Radcliffe Hospital, Oxford OX3 9DS, UK
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

Recombinant baculoviruses have established themselves as a favoured technology for the high-level expression of recombinant proteins. The construction of recombinant viruses, however, is a time consuming step that restricts consideration of the technology for high throughput developments. Here we use a targeted gene knockout technology to inactivate an essential viral gene that lies adjacent to the locus used for recombination. Viral DNA prepared from the knockout fails to initiate an infection unless rescued by recombination with a baculovirus transfer vector. Modified viral DNA allows 100% recombinant virus formation, obviates the need for further virus purification and offers an efficient means of mass parallel recombinant formation.


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