Basic ResearchSteatosis and liver cancer in transgenic mice expressing the structural and nonstructural proteins of hepatitis C virus☆,☆☆,★,★★
Section snippets
Production of transgenic mice
The plasmid pGEMAlbSVPA-B1(−) contains the murine albumin enhancer/promoter30 with a downstream SV40 intron/polyadenylation cassette. Complementary DNA (cDNA) corresponding to the full-length polyprotein-coding region (core to NS5B, nts 342–9378) or the structural protein-coding region only (core to E2/p7, nts 342–2771) of the genotype 1b, HCV-N strain of HCV31, 32 was inserted into this vector between newly engineered XhoI sites flanking the albumin enhancer/promoter and SV40 sequence,
Transgene expression in FL-N and S-N transgenic mice
Transgenic mice were designed to express either the full-length HCV-N open reading frame (FL-N) or the segment encoding the structural proteins only (S-N: core and the 2 envelope proteins, E1 and E2-p7; Figure 1).
Discussion
The HCV-transgenic mice that we have derived constitutively express low levels of RNA encoding the complete viral polyprotein, or higher levels of the RNA segment encoding only the structural proteins of the virus, under control of a liver-specific promoter. Protein expression was confirmed directly only in the mice expressing the structural region of the polyprotein and was insufficient for detection even with sensitive immunohistochemical techniques in mice with the full-length reading frame
Acknowledgements
The authors thank Annette Martin for critically reviewing the manuscript, Brian West for assistance in the interpretation of histopathology, Elbert Whorton for assistance with statistical analysis, and Li-Hua Ping and Geneviêve Inchauspé for generously providing HCV-specific antibodies.
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Address requests for reprints to: Stanley M. Lemon, M.D., Department of Microbiology and Immunology, The University of Texas Medical Branch, 301 University Boulevard, Galveston, Texas 77555-1019. e-mail: [email protected]; fax: (409) 772-3757.
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Dr. Lerat's present address: Institut de Génétique Moléculaire, CNRS, Montpellier, France. Dr. Gosert's present address: Institute for Medical Microbiology, University of Basel, Basel, Switzerland.
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Supported in part by the National Institute of Allergy and Infectious Diseases (U19-AI40035). H.L. was the recipient of a McLaughlin Fellowship from the University of Texas Medical Branch.
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Drs. Lerat and Honda contributed equally to this work.