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Efficient N-tailing of blunt DNA ends by Moloney murine leukemia virus reverse transcriptase
- Source :
- Scientific Reports
- Publication Year :
- 2016
-
Abstract
- Moloney murine leukemia virus reverse transcriptase (MMLV-RT) is a widely used enzyme for cDNA synthesis. Here we show that MMLV-RT has a strong template-independent polymerase activity using blunt DNA ends as substrate that generates 3′ overhangs of A, C, G, or T. Nucleotides were appended efficiently in the order A > G > T > C, and tail lengths varied from 4 to 5, 2 to 7, 2 to 4, and 2 to 3 for A, C, G, and T, respectively. The activity was so strong that nearly all our test DNA ends were appended with at least one A, C, G, or T. The N-tailing activity of MMLV-RT was enhanced in the presence of Mn2+, and the G-, C-, and T-tailing activities were further enhanced by dCMP, dGMP, and dAMP, respectively. This is the first report of an enzymatic activity that almost thoroughly appends two or more As, or one or more Cs, Gs, or Ts to the 3′ end of double-stranded DNA, which would enable exhaustive analysis of DNA samples. The N-tailing activity of MMLV-RT is potentially useful in many biotechnological applications.
- Subjects :
- 0301 basic medicine
DNA, Complementary
Article
03 medical and health sciences
Enzyme activator
chemistry.chemical_compound
Mice
Complementary DNA
Murine leukemia virus
Animals
Nucleotide
Polymerase
chemistry.chemical_classification
Multidisciplinary
030102 biochemistry & molecular biology
biology
RNA-Directed DNA Polymerase
DNA
Reverse Transcription
Templates, Genetic
biology.organism_classification
Molecular biology
Reverse transcriptase
Enzyme Activation
030104 developmental biology
Enzyme
chemistry
biology.protein
Moloney murine leukemia virus
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 7
- Database :
- OpenAIRE
- Journal :
- Scientific reports
- Accession number :
- edsair.doi.dedup.....1559847b32c2d6f9b3c2988e6d134bf6