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Journal of Virology, February 2001, p. 1284-1293, Vol. 75, No. 3
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.3.1284-1293.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Cloning and Sequencing of Defective Particles Derived from the Autonomous Parvovirus Minute Virus of Mice for the Construction of Vectors with Minimal cis-Acting Sequences

Nathalie Clément, Bernard Avalosse, Karim El Bakkouri, Thierry Velu, and Annick Brandenburger*

IRIBHN-IBMM, Université Libre de Bruxelles, B-6041 Gosselies, Belgium

Received 16 August 2000/Accepted 29 October 2000

The production of wild-type-free stocks of recombinant parvovirus minute virus of mice [MVM(p)] is difficult due to the presence of homologous sequences in vector and helper genomes that cannot easily be eliminated from the overlapping coding sequences. We have therefore cloned and sequenced spontaneously occurring defective particles of MVM(p) with very small genomes to identify the minimal cis-acting sequences required for DNA amplification and virus production. One of them has lost all capsid-coding sequences but is still able to replicate in permissive cells when nonstructural proteins are provided in trans by a helper plasmid. Vectors derived from this particle produce stocks with no detectable wild-type MVM after cotransfection with new, matched, helper plasmids that present no homology downstream from the transgene.

* Corresponding author. Mailing address: IRIBHN-IBMM, Université Libre de Bruxelles, rue des professeurs Jeener et Brachet, 12, B-6041 Gosselies, Belgium. Phone: (32 2) 650 98 31. Fax: (32 2) 650 98 20. E-mail: abranden{at}ulb.ac.be.

Journal of Virology, February 2001, p. 1284-1293, Vol. 75, No. 3
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.3.1284-1293.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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