Autographa californica nucleopolyhedrovirus (AcMNPV) has a large doublestrand DNA genome of approximately 134 kbp and comprises 156 open reading frames (ORFs). To elucidate DNA replication cascade of AcMNPV, we developed a novel baculovirus genome that can be maintained in Escherichia coli as a plasmid and can infect susceptible lepidopteran insect cells. This genome, named bAc-MK, contains a mini-F replicon and a kanamycin resistance marker. Using a convenient Tn7 transposon-based system, pPCS-S, which contains an ampicillin resistance gene, 56 single ORF-truncated mutants were generated by random insertion into bAc-MK genome. These single ORF-truncated mutants were independently transfected into Sf9 cells to verify viral replication. Interestingly, both lef-1 and p48 knockout mutants showed normal viral replication in infected cells, which are reported to essential for viral replication. These results suggest that these single ORF-truncated mutants are useful for elucidation of viral replication cascade.

• Xue Ying Tao(Department of Agricultural Biotechnology, Seoul National University)
• Jong Yul Roh(Department of Agricultural Biotechnology, Seoul National University)
• Jae Young Choi(Research Institute for Agriculture and Life Sciences, Seoul National University)
• Yong Wang(Department of Agricultural Biotechnology, Seoul National University)
• Qin Liu(Department of Agricultural Biotechnology, Seoul National University)
• Soo Dong Woo(College of Agriculture, Life and Environment Sciences, Chungbuk National University)
• Byung Rae Jin(College of Natural Resources and Life Science, Dong-A University)
• Yeon Ho Je(Department of Agricultural Biotechnology, Seoul National University)