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    Effect of in vitro transcription on cruciform stability.

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    Author
    Morales, Nydia M.; Cobourn, Scott D.; Muller, Uwe R.
    Abstract
    We have investigated the effect of in vitro transcription on cruciform stability. Replicative form DNA of 4X174 strain ins6240, containing a 48 bp synthetic palindrome in the J-F intercistronic region, was supercoiled in vitro to mean negative superhelical densities (ar) ranging from 0 to 0.15. The presence of cruciforms was probed by limited digestion with the single-strand specific nuclease BaIl3. The 48 bp palindrome was extruded at a mean a = - 0.05, but only after heating the DNA. An in vitro transcription reaction with E. coli RNA polymerase and [C-32P]UTP gave identical transcripts with heated or unheated template DNA. The synthetic cruciform was stable upon binding of the RNA polymerase to the template, but it was destabilized upon movement of the transcription complex along the template. Transcription of unheated templates did not result in cruciform formation. We propose that cruciform structures in supercoiled template DNAs present no hindrance to RNA polymerase, and thus have no detectable effect on transcription elongation in vitro. Originally published Nucleic Acids Research, Vol. 18, No. 9, May 1990
    URI
    http://hdl.handle.net/10342/3359
    Subject
     In vitro transcription; Cruciform stability 
    Date
    1990-05-11
    Citation:
    APA:
    Morales, Nydia M., & Cobourn, Scott D., & Muller, Uwe R.. (May 1990). Effect of in vitro transcription on cruciform stability.. Nucleic Acids Research, (18:9), p.2777-2782. Retrieved from http://hdl.handle.net/10342/3359

    Display/Hide MLA, Chicago and APA citation formats.

    MLA:
    Morales, Nydia M., and Cobourn, Scott D., and Muller, Uwe R.. "Effect of in vitro transcription on cruciform stability.". Nucleic Acids Research. 18:9. (2777-2782.), May 1990. August 08, 2022. http://hdl.handle.net/10342/3359.
    Chicago:
    Morales, Nydia M. and Cobourn, Scott D. and Muller, Uwe R., "Effect of in vitro transcription on cruciform stability.," Nucleic Acids Research 18, no. 9 (May 1990), http://hdl.handle.net/10342/3359 (accessed August 08, 2022).
    AMA:
    Morales, Nydia M., Cobourn, Scott D., Muller, Uwe R.. Effect of in vitro transcription on cruciform stability.. Nucleic Acids Research. May 1990; 18(9) 2777-2782. http://hdl.handle.net/10342/3359. Accessed August 08, 2022.
    Collections
    • Microbiology and Immunology
    Publisher
    East Carolina University

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