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Global Science Books

http://globalsciencebooks.com

Advances in Gene, Molecular and Cell Therapy

Volume 1 Number 2 2007

Michèle Amouyal (France) Use of Histones to Increase the Frequency of Recombinant Plasmid
Formation during Molecular Cloning (pp 192-197)

CNRS-Interactions-à-Distance. E-mail: michele.amouyal@mail.com

ABSTRACT
Research Note: By compacting DNA in eukaryotes, histones bring closer two distant regions on DNA.
This is likely to facilitate the work of transcriptional enhancers at a long distance in gene regulation
(Amouyal 1991). Since the classical strategy for plasmid engineering involves a step where the
recombinant fragment is circularized, this has prompted the author to test this hypothesis by plasmid
engineering. In fact, the unfractionated mixture of calf thymus histones facilitates the in vitro ligation of
two fragments, resulting in an increased number of transformed E. coli cells under the appropriate
conditions for about 7 and 12 kb large recombinant plasmids. This property can be exploited for
plasmid engineering, when inserting a DNA fragment into a plasmid vector becomes increasingly
difficult as the recombinants get larger. The same effect was obtained with calf thymus histone H2B by
itself, but was less marked with the linker histone H1.