Flexible Genetic Engineering Using RecA Protein
By: Lance J. Ferrin2
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The analysis or manipulation of small segments of DNA in plasmids has long been routine, but standard methods fail or are
very time consuming when applied to long segments or complex mixtures of DNA. At present, RecA protein-based techniques usually
have been used for making defined changes in large DNA constructs or for precise physical mapping of large regions of genomic
DNA from several organisms. The techniques offer a rapid and versatile “cut and paste” approach to several other problems
in molecular biology, and their use will probably increase as the goals of experiments with DNA and chromosomes evolve in
sophistication. For example, they can now be used to analyze rearranged DNA in individual tumors or genetic deletions in patients
with inherited or acquired syndromes. A new variation involving sequencespecific ligation may also eventually be used to rapidly
screen libraries, label DNA fragments, or amplify large segments of DNA (1).
Affiliation(s): (2) Division of Gastroenterology, Department of Medicine, University of Minnesota, Minneapolis, MN
Book Title: DNA Repair Protocols: Prokaryotic Systems
Series: Methods in Molecular Biology | Volume: 152 | Pub. Date: Jul-07-2000 | Page Range: 135-147 | DOI: 10.1385/1-59259-068-3:135
Subject: Genetics/Genomics
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