Extended minus-strand DNA as template for R-U5-mediated second-strand transfer in recombinational rescue of primer binding site-modified retroviral vectors.
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Extended minus-strand DNA as template for R-U5-mediated second-strand transfer in recombinational rescue of primer binding site-modified retroviral vectors. / Mikkelsen, J G; Lund, Anders Henrik; Dybkaer, K; Duch, M; Pedersen, F S.
In: Journal of Virology, Vol. 72, No. 3, 1998, p. 2519-25.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Extended minus-strand DNA as template for R-U5-mediated second-strand transfer in recombinational rescue of primer binding site-modified retroviral vectors.
AU - Mikkelsen, J G
AU - Lund, Anders Henrik
AU - Dybkaer, K
AU - Duch, M
AU - Pedersen, F S
N1 - Keywords: 3T3 Cells; Animals; Base Sequence; Binding Sites; Cell Transformation, Viral; DNA Primers; DNA, Single-Stranded; DNA, Viral; Genetic Vectors; Leukemia Virus, Murine; Mice; Molecular Sequence Data; Recombination, Genetic; Repetitive Sequences, Nucleic Acid; Ribonucleoprotein, U5 Small Nuclear; Templates, Genetic; Virus Replication
PY - 1998
Y1 - 1998
N2 - We have previously demonstrated recombinational rescue of primer binding site (PBS)-impaired Akv murine leukemia virus-based vectors involving initial priming on endogenous viral sequences and template switching during cDNA synthesis to obtain PBS complementarity in second-strand transfer of reverse transcription (Mikkelsen et al., J. Virol. 70:1439-1447, 1996). By use of the same forced recombination system, we have now found recombinant proviruses of different structures, suggesting that PBS knockout vectors may be rescued through initial priming on endogenous virus RNA, read-through of the mutated PBS during minus-strand synthesis, and subsequent second-strand transfer mediated by the R-U5 complementarity of the plus strand and the extended minus-strand DNA acceptor template. Mechanisms for R-U5-mediated second-strand transfer and its possible role in retrovirus replication and evolution are discussed.
AB - We have previously demonstrated recombinational rescue of primer binding site (PBS)-impaired Akv murine leukemia virus-based vectors involving initial priming on endogenous viral sequences and template switching during cDNA synthesis to obtain PBS complementarity in second-strand transfer of reverse transcription (Mikkelsen et al., J. Virol. 70:1439-1447, 1996). By use of the same forced recombination system, we have now found recombinant proviruses of different structures, suggesting that PBS knockout vectors may be rescued through initial priming on endogenous virus RNA, read-through of the mutated PBS during minus-strand synthesis, and subsequent second-strand transfer mediated by the R-U5 complementarity of the plus strand and the extended minus-strand DNA acceptor template. Mechanisms for R-U5-mediated second-strand transfer and its possible role in retrovirus replication and evolution are discussed.
M3 - Journal article
C2 - 9499117
VL - 72
SP - 2519
EP - 2525
JO - Journal of Virology
JF - Journal of Virology
SN - 0022-538X
IS - 3
ER -
ID: 5016576