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Journal of Virology, June 2002, p. 6393-6397, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.6393-6397.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of the Lymphocytic Choriomeningitis Virus (LCMV) Proteins Required To Rescue LCMV RNA Analogs into LCMV-Like Particles{dagger}

Ki Jeong Lee,,{ddagger} Mar Perez, Daniel D. Pinschewer, and Juan Carlos de la Torre*

Department of Neuropharmacology, The Scripps Research Institute, La Jolla, California 92037

Received 2 January 2002/ Accepted 19 March 2002

We have used a reverse genetic approach to identify the viral proteins required for packaging and assembly of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV). Plasmids encoding individual LCMV proteins under the control of an RNA polymerase II promoter were cotransfected with a plasmid containing an LCMV minigenome (MG). Intracellular synthesis of the LCMV MG was driven by T7 RNA polymerase whose expression was also mediated by a Pol II promoter. The supernatant from transfected cells was passaged onto fresh cells that were subsequently infected with LCMV to provide the minimal viral trans-acting factors, NP and L, that are required for LCMV MG RNA replication and expression. Reconstitution of LCMV-specific packaging and passage was detected by expression of the chloramphenicol acetyl transferase (CAT) reporter gene present in the MG. NP and L did not direct detectable levels of MG passage. Addition of Z and GP resulted in high levels of passage of CAT activity, which could be prevented by LCMV neutralizing antibodies. Passage of LCMV MG was inhibited by omission of either GP or Z.

* Corresponding author. Mailing address: Department of Neuropharmacology, IMM-6, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037. Phone: (858) 784-9562. Fax: (858) 784-9981. E-mail: juanct{at}scripps.edu.

{dagger} This is publication 14640-NP from The Scripps Research Institute.

{ddagger} Present address: Department of Molecular Microbiology and Immunology, University of Southern California School of Medicine, HMR 401, Los Angeles, CA 90033.

Journal of Virology, June 2002, p. 6393-6397, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.6393-6397.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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