| dc.description.abstract | The Sudden Acute Respiratory Syndrome Coronavirus (SARS-CoV) caused a severe atypical pneumonia in humans with close to a 10% mortality rate, and caused a global outbreak in 2003. To better understand the pathogenesis of coronavirus pneumovirulence, a related coronavirus model is being developed, Murine Hepatitis Virus 1 (MHV-1). A complete reverse genetic system for MHV-1 is in the process of being completed. The reverse genetic system uses 7 cDNA fragments that cover the entire MHV-1 genome. These fragments are recovered from reverse transcribed viral RNA, and unique restriction sites were added to flank each fragment in order to allow systematic assembly of the complete genomic cDNA. The genomic cDNA can be transcribed, and the recovered RNA can be transfected into susceptible mammalian cells to rescue virus. The transfection process is currently being optimized by investigating the benefits of different electroporation techniques and different overlay cell confluences on amount of infectious centers produced from each attempt of viral recovery. Once the reverse genetic system is complete and tested the system will be used to generate mutant virus, impossible to make with out the reverse genetic system. | en |
