The reverse genetics system is a very powerful tool for analyzing the molecular mechanisms of viral propagation and pathogenesis. However, full-length genome plasmid construction is highly time-consuming and laborious, and undesired mutations may be introduced by Escherichia coli. This study shows a very rapid E. coli-free method of full-genome construction using the mumps virus as an example. This method was able to reduce dramatically the time for full-genome construction, which was used very efficiently for virus rescue, from several days or more to ~2 days, with a similar accuracy and yield to the conventional method using E. coli/plasmid.
Keywords: Escherichia coli-free; OriCiro cell-free cloning system; PCR; mumps virus; reverse genetics.
© 2022 The Authors. Microbiology and Immunology published by The Societies and John Wiley & Sons Australia, Ltd.