Abutilon mosaic geminivirus proteins expressed and phosphorylated in Escherichia coli

In order to get more detailed information on single proteins involved in th e life cycle of Abutilon mosaic geminivirus, special conditions were establ ished allowing expression of the proteins encoded by different viral open r eading frames (AC2, AC3, AV1, BC1, and BV1) in Escherichia coli by use of t he pET expression vector system. These include cotransformation of a tRNA s upplementation plasmid to overcome codon usage problems in the case of AC2. Protocols were developed to purify the virus proteins for production of an tisera. Phosphorylation of AbMV proteins AC3, AV1, BC1, and BV1 in E. coli could be shown by autoradiographic detection of stably incorporated inorgan ic (PO43-)-P-32.