Expression of a foreign gene by recombinant canine distemper virus recovered from cloned DNAs

Christopher L. Parks, Hai Ping Wang, Gerald R. Kovacs, Nikos Vasilakis, Jacek Kowalski, Rebecca M. Nowak, Robert A. Lerch, Pramila Walpita, Mohinderjit S. Sidhu, Stephen A. Udem

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

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°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.

Original languageEnglish (US)
Pages (from-to)131-147
Number of pages17
JournalVirus Research
Volume83
Issue number1-2
DOIs
StatePublished - Feb 26 2002
Externally publishedYes

Fingerprint

Canine Distemper Virus
Viruses
DNA
Genes
Luciferases
Sequence Analysis
Vero Cells
Reporter Genes
Shock
Plasmids
Vaccines
Complementary DNA
Clone Cells
Cell Culture Techniques
Hot Temperature

Keywords

  • cDNA rescue
  • Foreign gene expression
  • Recombinant canine distemper virus

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

Cite this

Parks, C. L., Wang, H. P., Kovacs, G. R., Vasilakis, N., Kowalski, J., Nowak, R. M., ... Udem, S. A. (2002). Expression of a foreign gene by recombinant canine distemper virus recovered from cloned DNAs. Virus Research, 83(1-2), 131-147. https://doi.org/10.1016/S0168-1702(01)00430-0

Expression of a foreign gene by recombinant canine distemper virus recovered from cloned DNAs. / Parks, Christopher L.; Wang, Hai Ping; Kovacs, Gerald R.; Vasilakis, Nikos; Kowalski, Jacek; Nowak, Rebecca M.; Lerch, Robert A.; Walpita, Pramila; Sidhu, Mohinderjit S.; Udem, Stephen A.

In: Virus Research, Vol. 83, No. 1-2, 26.02.2002, p. 131-147.

Research output: Contribution to journalArticle

Parks, CL, Wang, HP, Kovacs, GR, Vasilakis, N, Kowalski, J, Nowak, RM, Lerch, RA, Walpita, P, Sidhu, MS & Udem, SA 2002, 'Expression of a foreign gene by recombinant canine distemper virus recovered from cloned DNAs', Virus Research, vol. 83, no. 1-2, pp. 131-147. https://doi.org/10.1016/S0168-1702(01)00430-0
Parks, Christopher L. ; Wang, Hai Ping ; Kovacs, Gerald R. ; Vasilakis, Nikos ; Kowalski, Jacek ; Nowak, Rebecca M. ; Lerch, Robert A. ; Walpita, Pramila ; Sidhu, Mohinderjit S. ; Udem, Stephen A. / Expression of a foreign gene by recombinant canine distemper virus recovered from cloned DNAs. In: Virus Research. 2002 ; Vol. 83, No. 1-2. pp. 131-147.
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