Minimum origin requirements for linear duplex AAV DNA replication in vitro.
We have investigated the minimal requirements for a functional origin of DNA replication in an in vitro assay which requires the adeno-associated virus (AAV) Rep 68/78 protein. When a linear duplex template was used, initiation of one round of DNA replication was achieved when nucleotides 1-56 of the AAV inverted terminal repeat (in the flop orientation) were present at the terminus of the template. A terminal resolution site was not required. Deletion of an additional 11 nucleotides from the terminal repeat blocked the reaction.
Ward
P
P
Department of Microbiology, W. R. Hearst Microbiology Research Center, Cornell University Medical College, New York, New York 10021, USA.
Berns
K I
KI
D004261:DNA Replication; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism*; D016679:Genome, Viral; D008957:Models, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D018741:Replication Origin; D015183:Restriction Mapping; D017384:Sequence Deletion; D013698:Templates, Genetic