Expression of a foreign gene by recombinant canine distemper virus recovered from cloned DNAs
| Autor: | Christopher L. Parks, Stephen A. Udem, Gerald R. Kovacs, Robert A. Lerch, Rebecca M. Nowak, Hai Ping Wang, Pramila Walpita, Nikos Vasilakis, Jacek Kowalski, Mohinderjit S. Sidhu |
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| Rok vydání: | 2002 |
| Předmět: |
Cancer Research
viruses Genetic Vectors Molecular Sequence Data Gene Expression Biology Virus law.invention Cell Line Plasmid Dogs law Genes Reporter Virology Cricetinae Chlorocebus aethiops medicine Tumor Cells Cultured Animals Humans Luciferase Cloning Molecular Luciferases Distemper Virus Canine Vero Cells Recombination Genetic Reporter gene Base Sequence Canine distemper Transfection DNA-Directed RNA Polymerases Nucleocapsid Proteins medicine.disease Molecular biology Infectious Diseases DNA Viral Recombinant DNA Vero cell Replicon |
| Zdroj: | Virus research. 83(1-2) |
| ISSN: | 0168-1702 |
| Popis: | A canine distemper virus (CDV) genomic cDNA clone and expression plasmids required to establish a CDV rescue system were generated from a laboratory-adapted strain of the Onderstepoort vaccine virus. In addition, a CDV minireplicon was prepared and used in transient expression studies performed to identify optimal virus rescue conditions. Results from the transient expression experiments indicated that minireplicon-encoded reporter gene activity was increased when transfected cell cultures were maintained at 32 rather than 37 degrees C, and when the cellular stress response was induced by heat shock. Applying these findings to rescue of recombinant CDV (rCDV) resulted in efficient recovery of virus after transfected HEp2 or A549 cells were co-cultured with Vero cell monolayers. Nucleotide sequence determination and analysis of restriction site polymorphisms confirmed that rescued virus was rCDV. A rCDV strain also was engineered that contained the luciferase gene inserted between the P and M genes; this virus directed high levels of luciferase expression in infected cells. |
| Databáze: | OpenAIRE |
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