Abstract
E. coli DNA polymerase I (EC 2.7.7.7) can engage in either DNA- or RNA-directed DNA synthesis with hybrid templates. The choice of the strand to be transcribed depends primarily on the relative lengths of the two strands of the hybrid, the longer strand serving as the template and the shorter as the primer. If a polynucleotide is reduced in size by exposure to an endonuclease before being hybridized to the complementary strand, the template efficiency of the latter increases several-fold. Under properly selected conditions, highly efficient reverse transcription of the all-ribonucleotide template-primers poly(A).oligo(U), poly(C).oligo(I), and poly(I).oligo(C) can be achieved. "f1 RNA," the RNA strand of an f1 DNA.RNA hybrid, can also serve as template for reverse transcription either after "nicking" of the hybrid with DNase, or after separation from the DNA strand and priming by DNase-treated f1 DNA.
MeSH Terms
Adenine Nucleotides
Adenosine Triphosphate/metabolism
Animals
DNA, Bacterial/biosynthesis
Deoxyribonucleases/metabolism
Escherichia coli/enzymology
Kidney/enzymology
Magnesium
Manganese
Nucleic Acid Hybridization
Poly I-C
Polynucleotides
Potassium Chloride
RNA-Directed DNA Polymerase/metabolism
Ribonucleases/metabolism
Sheep
Sodium Chloride
Templates, Genetic
Thymine Nucleotides/metabolism
Tritium
Uracil Nucleotides
Chemicals
Adenine Nucleotides
DNA, Bacterial
Polynucleotides
Thymine Nucleotides
Uracil Nucleotides
Tritium
Manganese
Sodium Chloride
Potassium Chloride
Adenosine Triphosphate
RNA-Directed DNA Polymerase
Deoxyribonucleases
Ribonucleases
Magnesium
Poly I-C
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Karkas J D
References (22)
22 references, click to expand
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