Linked Life Data

18770555

Source:http://linkedlifedata.com/resource/pubmed/id/18770555

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2008-9-4
pubmed:abstractText
This unit contains protocols for evaluation of replication functionality of papillomavirus genomes or subgenomic fragments. Replication is measured after transient cotransfection of the genome (or subgenomic fragment) with expression vectors encoding the viral E1 and E2 proteins. Input DNA is methylated at the adenine of GATC sequences by propagation in E. coli. DNA that replicates in mammalian cells will lose the adenine methylation and become DpnI-resistant, while residual methylated input DNA will remain DpnI-sensitive. After transfection, DNA extraction, and DpnI digestion, replicated DNA can be detected by Southern blotting as a full-length plasmid, since it is resistant to digestion. This assay can be used to map the genomic location of a functional origin or to evaluate replication activity of mutations in either the origin DNA sequences or the E1 or E2 proteins.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
1934-8533
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
Chapter 14
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
Unit 14B.1
pubmed:meshHeading
pubmed:year
2005
pubmed:articleTitle
Cell culture assay for transient replication of human and animal papillomaviruses.
pubmed:affiliation
Texas A&M University System Health Science Center, College Station, Texas, USA.
pubmed:publicationType
Journal Article