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Titolo:
Polymerase activities and RNA structures in the atomic force microscope
Autore:
Hansma, HG; Golan, R; Hsieh, W; Daubendiek, SL; Kool, ET;
Indirizzi:
Univ Calif Santa Barbara, Dept Phys, Santa Barbara, CA 93106 USA Univ Calif Santa Barbara Santa Barbara CA USA 93106 Barbara, CA 93106 USA Univ Rochester, Dept Chem, Rochester, NY 14627 USA Univ Rochester Rochester NY USA 14627 Dept Chem, Rochester, NY 14627 USA
Titolo Testata:
JOURNAL OF STRUCTURAL BIOLOGY
fascicolo: 3, volume: 127, anno: 1999,
pagine: 240 - 247
SICI:
1047-8477(199910)127:3<240:PAARSI>2.0.ZU;2-K
Fonte:
ISI
Lingua:
ENG
Soggetto:
DEPENDENT PROTEIN-KINASE; CIRCULAR OLIGONUCLEOTIDES; ROLLING TRANSCRIPTION; SINGLE MOLECULES; CATALYTIC RNAS; DNA; VISUALIZATION; COMPLEXES; GENERATION; DISSECTION;
Keywords:
AFM; AMV reverse transcriptase; DNA rolling circle; enzyme assay; replication; ribozyme; Sequenase; scanning force microscope; SFM; scanning probe microscopy; SPM; Taq polymerase; transcription;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
49
Recensione:
Indirizzi per estratti:
Indirizzo: Hansma, HG Univ Calif Santa Barbara, Dept Phys, Santa Barbara, CA 93106 USA Univ Calif Santa Barbara Santa Barbara CA USA 93106 93106 USA
Citazione:
H.G. Hansma et al., "Polymerase activities and RNA structures in the atomic force microscope", J STRUCT B, 127(3), 1999, pp. 240-247

Abstract

The structures of the reaction products are the basis for novel polymeraseassays using the atomic force microscope (AFM). Polymerases are the enzymes involved in transcription and replication of DNA Rapid semiquantitative estimates of the activity of DNA polymerases such as Sequenase, Tag polymerase, and AMV reverse transcriptase and RNA polymerases (RNAP) such as Escherichia coli RNAP were obtained from AFM images of the nucleic acids after polymerase reactions. DNA polymerases were assayed via replication of the single-stranded phi X-174 virion. RNAP was assayed via transcription, using a rolling circle DNA template that produces long strands of RNA. In some cases, AFM was better than agarose gel electrophoresis for assaying DNA polymerase activity, since aggregation prevented the DNA from entering the agarosegel. Extended molecules of single-stranded RNA synthesized with the rolling circle DNA template showed varied conformations and degrees of stretching. Some structural differences were observed between two RNAs-a ribozyme concatamer and an RNA with 90% purines. (C) 1999 Academic Press.

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Documento generato il 05/04/20 alle ore 19:22:18