TAILIEUCHUNG - Báo cáo khoa học: "Targeted RNA Recombination of the Membrane Nucleocapsid Protein Genes between Mouse Hepatitis Virus Bovine Coronavirus"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Targeted RNA Recombination of the Membrane Nucleocapsid Protein Genes between Mouse Hepatitis Virus Bovine Coronavirus | J. Vet. Sci. 2001 2 3 149-157 IDDHBBHHHBH Veterinary Science Targeted RNA Recombination of the Membrane and Nuclrocapsid Protein Genes between Mouse Hepatitis Virus and Bovine Coronavirus Gabriella Koljesar5 and Dongwan Yoo Department of Pathobiology Ontario Veterinary College University of Guelph Guelph Ontario NIG 2W1 Canada ABSTRACT The targeted RNA recombination was attempted to substitute the membrane M protein gene and part of the nucleocapsid N protein gene OỈ mouse hepatitis virus with the corresponding sequences from bovine coronavirus. Using a defective interfering DI RNA-like cDNA construct derived from pMH54 090 nucleotides representing the entire M gene and the 5 most 915 nucleotides of the N gene of the mouse hepatitis virus Albany 4 mutant were attempted to be replaced. Upon infection of cells with Albany 4 followed by transfection with synthetic RNA transcribed from the DI-like cDNA construct recombinant mouse hepatitis viruses as the large plaque forming phenotype were isolated by plaque assays at the non-permissive temperature of 39 ĨU By RT-PCR and sequencing those large plaque phenotypes were confirmed to have contained the thermostable phenotype marker derived from the transfected RNA demonstrating that recombination occurred between the Albany 4 genomic RNA and the in vitro RNA transcripts. Further analysis erf the recombinant viruses indicated that the recombination had taken place within the region of 222 nucleotides between positions 916 and 1137 of the N gene. This is the region immediately downstream of the replacement sequence and the start of the temperature resistant phenotype marker. The results suggest that the M and part of the N genes of bovine coronavirus may not be able to complement the function of those of Corresponding author address Dr. Dongwan Yoo Department of Pathobiology Ontario Veterinary College University of Guelph Guelph Ontario NIG 2W1 Canada. Email dyoo@ Phone 519-824-4120 ext 4729 Fax 519-767-0809 s GK .

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