Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1989-02-24
1992-01-07
Schwartz, Richard A.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
536 27, 435183, 435822, C12P 1934, C07H 2100, C12N 900
Patent
active
050791516
ABSTRACT:
msDNA is a peculiar molecule consisting of a branched RNA linked to single-stranded DNA via a 2', 5' phosphodiester bond. A cell-free system, utilizing cells permeablized with phenethyl, alcohol, was established to study the synthesis of msDNA in Myxococcus xanthus. Permeablized cell labeled with [.alpha.-.sup.32 P]dCTP in the presence of ddGTP, ddATP, or ddTTP, produce a band that migrates at the same position as the full-sized msDNA in an acrylamide gel. However, when this band is treated with ribonuclease A prior to gel electrophoresis, it resulted in many bands of different sizes. This indicates that during the labeling, intermediates are produced in which single-stranded DNAs of various lengths are associated with a compensatory length of RNA such that the total length for each intermediate is identical. These results provide evidence for the previously proposed model in which msDNA is synthesized by reverse transcriptase using a folded RNA precursor as a primer as well as a template. Furthermore, it was found that there is a precise coupling mechanism of reverse transcriptase and ribonuclease H.
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Inouye Masayori
Inouye Sumiko
Lampson Bert C.
Mosher Mary E.
Schwartz Richard A.
University of Medicine and Dentistry of New Jersey
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