Cloning of adeno-associated virus into pBR322: rescue of intact virus from the recombinant plasmid in human cells.
RJ Samulski, KI Berns, M Tan… - Proceedings of the …, 1982 - National Acad Sciences
RJ Samulski, KI Berns, M Tan, N Muzyczka
Proceedings of the National Academy of Sciences, 1982•National Acad SciencesWe have cloned intact duplex adeno-associated virus (AAV) DNA into the bacterial plasmid
pBR322. The AAV genome could be rescued from the recombinant plasmid by transfection
of the plasmid DNA into human cells with adenovirus 5 as helper. The efficiency of rescue
from the plasmid was sufficiently high to produce yields of AAV DNA comparable to those
observed after transfection with equal amounts of purified virion DNA. Thus, the recombinant
plasmid itself may be a model for studying the rescue of a latent AAV viral infection. In …
pBR322. The AAV genome could be rescued from the recombinant plasmid by transfection
of the plasmid DNA into human cells with adenovirus 5 as helper. The efficiency of rescue
from the plasmid was sufficiently high to produce yields of AAV DNA comparable to those
observed after transfection with equal amounts of purified virion DNA. Thus, the recombinant
plasmid itself may be a model for studying the rescue of a latent AAV viral infection. In …
We have cloned intact duplex adeno-associated virus (AAV) DNA into the bacterial plasmid pBR322. The AAV genome could be rescued from the recombinant plasmid by transfection of the plasmid DNA into human cells with adenovirus 5 as helper. The efficiency of rescue from the plasmid was sufficiently high to produce yields of AAV DNA comparable to those observed after transfection with equal amounts of purified virion DNA. Thus, the recombinant plasmid itself may be a model for studying the rescue of a latent AAV viral infection. In addition, the efficient rescue of viable AAV from the recombinant plasmid should facilitate the genetic analysis of AAV. Finally, the results of an analysis of the DNA from rescued virions indicate that an inversion of the AAV terminal sequences occurred during replication.
National Acad Sciences