High-level expression of active HIV-1 integrase from a synthetic gene in human cells
| Autor: | Paul Proost, Peter Cherepanov, Wim Pluymers, Anje Claeys, Zeger Debyser, Erik De Clercq |
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| Rok vydání: | 2000 |
| Předmět: |
Genes
Viral Genetic Vectors Molecular Sequence Data DNA Recombinant Gene Expression HIV Integrase Transfection Biochemistry Defective virus Cell Line Viral vector Enzyme Stability Genetics Humans Amino Acid Sequence Viral Accessory Proteins Molecular Biology Gene DNA Primers Cell Nucleus Base Sequence biology Genetic Complementation Test HEK 293 cells Defective Viruses Molecular biology Recombinant Proteins Integrase Cell culture HIV-1 Mutagenesis Site-Directed biology.protein HeLa Cells Biotechnology |
| Zdroj: | The FASEB Journal. 14:1389-1399 |
| ISSN: | 1530-6860 |
| Popis: | A synthetic gene encoding for HIV-1 integrase was designed to circumvent the intrinsic instability and the repressor elements present in the wild-type gene. High-level expression of HIV-1 integrase was obtained in various human cell lines independently of viral accessory proteins. A human 293T cell line was selected that stably expresses HIV-1 integrase and has growth kinetics comparable to the parental cell line. The enzyme was localized in the nucleus and remained stably associated with the chromosomes during mitosis. Lentiviral vector particles carrying the inactivating D64V mutation in the integrase gene were capable of stably transducing 293T cells when complemented in the producer cells with integrase expressed from the synthetic gene. When the cell line that stably expresses integrase was infected with the defective viral particles, complementation of integrase activity was detected as well. Expression of active HIV-1 integrase in human cells will facilitate the study of the interplay between host and viral factors during integration. |
| Databáze: | OpenAIRE |
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