transform bacteria with linear DNA?

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Wed Sep 27 17:28:44 EST 1995


Pat Martin (pkmartin at netcom.com) writes:

| after reading about the number of experimental options the yeast
| community has at their disposal for the purpose of studying gene
| function, a question arose about bacteria.  budding yeast (and perhaps
| fission yeast as well) can be mutagenized by transformation with
| oligonucleotides and genes can be disrupted with the appropriately
| designed PCR fragments.  can anyone point me to a reference as to why
| this isnt done in bacteria?

It is! You should have a look at this article:

@article{Winans1985,
author = "S. C. Winans
     and S. J. Elledge
     and J. H. Krueger
     and G. C. Walker",
title = "Site-directed insertion and deletion mutagenesis
with cloned fragments in {{\em Escherichia coli}}",
journal = "J. Bacteriol.",
volume = "161",
pages = "1219-1221",
year = "1985"}

abstract:

A mutation of a cloned gene that has been made by introducing a
transposon or some other selectable genetic determinant can be crossed
into the gene's original replicon by linearizing the cloned DNA and
transforming a recB recC sbcB mutant. A number of applications of this
method are described with genes of either chromosomal or plasmid origin.

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