Abstract:
:We present a method for studying multiple retroviral integration events into a small DNA target in vivo. Episomal simian virus 40 (SV40) genomes established by infection of CV-1 cells served as integration targets during subsequent infection with murine leukemia virus (MLV). Using a PCR-based assay for the abundance and distribution of integration events, nonrandom integration of MLV DNA into SV40 DNA is detectable as early as 4 hr and reaches a maximum level by 8 hr after MLV infection. The level of integration but not the distribution of integration sites is sensitive to the stage in the SV40 life cycle at which MLV infection is performed. Using a temperature-sensitive tumor (T) antigen mutant SV40 strain, we observed that active replication of the target DNA is not required for efficient integration in vivo. The distribution of integration sites in vivo is closely approximately by in vitro reactions with isolated SV40 minichromosomes as integration targets. However, the degree of bias between the most and least favored sites is greater in vivo than in vitro.
journal_name
Proc Natl Acad Sci U S Aauthors
Pryciak PM,Müller HP,Varmus HEdoi
10.1073/pnas.89.19.9237keywords:
subject
Has Abstractpub_date
1992-10-01 00:00:00pages
9237-41issue
19eissn
0027-8424issn
1091-6490journal_volume
89pub_type
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journal_title:Proceedings of the National Academy of Sciences of the United States of America
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journal_title:Proceedings of the National Academy of Sciences of the United States of America
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更新日期:2016-11-08 00:00:00