Literature DB >> 28660237

Analysis of Replicative Intermediates of Adeno-associated Virus through Hirt Extraction and Southern Blotting.

Martino Bardelli1, Francisco Zarate-Perez2, Leticia Agundez1, Nelly Jolinon1, R Michael Linden1, Carlos R Escalante2, Els Henckaerts1.   

Abstract

Adeno-associated virus (AAV) is a small single-stranded DNA virus that requires the presence of a helper virus, such as adenovirus or herpes virus, to efficiently replicate its genome. AAV DNA is replicated by a rolling-hairpin mechanism (Ward, 2006), and during replication several DNA intermediates can be detected. This detailed protocol describes how to analyze the AAV DNA intermediates formed during AAV replication using a modified Hirt extract (Hirt, 1967) procedure and Southern blotting (Southern, 1975).

Entities:  

Keywords:  AAV; Adeno-associated virus; DNA replication; Replicative intermediates; Southern blot

Year:  2017        PMID： 28660237      PMCID： PMC5484404          DOI： 10.21769/BioProtoc.2271

Source DB:  PubMed          Journal:  Bio Protoc        ISSN： 2331-8325


  9 in total

1.  Detection of specific sequences among DNA fragments separated by gel electrophoresis.

Authors:  E M Southern
Journal:  J Mol Biol       Date:  1975-11-05       Impact factor: 5.469

2.  Selective extraction of polyoma DNA from infected mouse cell cultures.

Authors:  B Hirt
Journal:  J Mol Biol       Date:  1967-06-14       Impact factor: 5.469

3.  Evidence for a single-stranded adenovirus-associated virus genome: formation of a DNA density hybrid on release of viral DNA.

Authors:  J A Rose; K I Berns; M D Hoggan; F J Koczot
Journal:  Proc Natl Acad Sci U S A       Date:  1969-11       Impact factor: 11.205

4.  Manipulation of adenovirus vectors.

Authors:  F L Graham; L Prevec
Journal:  Methods Mol Biol       Date:  1991

5.  The AAV origin binding protein Rep68 is an ATP-dependent site-specific endonuclease with DNA helicase activity.

Authors:  D S Im; N Muzyczka
Journal:  Cell       Date:  1990-05-04       Impact factor: 41.582

6.  Cloning of adeno-associated virus into pBR322: rescue of intact virus from the recombinant plasmid in human cells.

Authors:  R J Samulski; K I Berns; M Tan; N Muzyczka
Journal:  Proc Natl Acad Sci U S A       Date:  1982-03       Impact factor: 11.205

7.  Concatemers of alternating plus and minus strands are intermediates in adenovirus-associated virus DNA synthesis.

Authors:  S E Straus; E D Sebring; J A Rose
Journal:  Proc Natl Acad Sci U S A       Date:  1976-03       Impact factor: 11.205

8.  Cloning of infectious adeno-associated virus genomes in bacterial plasmids.

Authors:  C A Laughlin; J D Tratschin; H Coon; B J Carter
Journal:  Gene       Date:  1983-07       Impact factor: 3.688

9.  Identification of a Functionally Relevant Adeno-Associated Virus Rep68 Oligomeric Interface.

Authors:  Martino Bardelli; Francisco Zárate-Pérez; Leticia Agúndez; R Michael Linden; Carlos R Escalante; Els Henckaerts
Journal:  J Virol       Date:  2016-07-11       Impact factor: 5.103
  9 in total

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