Kanamycin resistance gene

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Tue Feb 23 13:58:38 EST 1993

In article <1993Feb23.004359.23960 at magnus.acs.ohio-state.edu>
 gchacko at magnus.acs.ohio-state.edu (George W Chacko) writes:

>In article <C2v9Ms.487 at ccu.umanitoba.ca>
 hamel at ccu.umanitoba.ca (Andre Hamel) writes:
>>Perhaps publications by V.N.Iyer of Carleton University in Ottawa, Canada may
>>be of help (particularly some that come to mind by Selvaraj of Iyer's lab,
>>back sometime around 1982 to 1987). There were vectors that were part of 
>>lineages of vectors that come to mind, so you may have to look at their
>>plasmid diagrams carefully. Sorry I don't have more specific details on
>>hand, but this was five years ago since I was last "intimate" with these
>>particular vectors (I modified some myself but they wouldn't be of use to
>>you due to too few remaining rest. endo. cleavage sites).
>As I remember, there were a series of Tn5 containing plasmids that
>came out of that lab. I cloned kanamycin resistance into pUC19's
>HindIII site (It's easy. Just select on kanamycin plates) but have no
>idea where that plasmid is now. I also remember a plasmid from John
>Beringer that was a Tn5 delivery vector that you might want to look

I have made a pUC derivative which has a kanamycin resistance cassette you
may find useful. On the plasmid pNH-Kan/oriT, there is a 1.4 kb kanamycin
resistance gene that can be easily subcloned onto any plasmid.

The reference is:

AUTHOR = "P. N. Hengen
        and V. N. Iyer",
TITLE = "DNA cassettes containing the origin of transfer (oriT) of two
        broad-host-range transfer systems",
JOURNAL = "Biotechniques",
YEAR = "1992",
VOLUME = "13",
PAGES = "57-62"}


Plasmid constructs are described which carry retrievable DNA cassettes
containing the origin of transfer region (oriT) from two broad-host-range
plasmids. Restriction of these high copy number plasmids with any one of a
variety of enzymes yields a linear DNA fragment of convenient size
containing the oriT region of either pCU1 or RK2. This DNA can be
ligated into any vector or recombinant plasmid containing a compatible
enzyme site and can be easily identified by size on an agarose gel.
Any plasmid can therefore be mobilized using a number of helper strains or
conjugative plasmids derived from the parental plasmids. In addition, the
cassettes can be used for a variety of genetic manipulations including
"selectable" linker mutagenesis.

The plasmids described in this paper have been deposited with the
American Type Culture Collection (ATCC). pUC1813-NoriT is number
77276 and pNH-Kan/oriT is number 77277. They are available for a
nominal fee. You can get more information by contacting:

American Type Culture Collection
12301 Parklawn Drive
Rockville, Maryland 20852 USA
phone (301) 231-5585

You can also access the ATCC catalog via modem. The number is
1-800-647-4710 use vt100 2400b 8,1,N
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Paul N. Hengen
National Cancer Institute
Frederick Cancer Research and Development Center
Frederick, Maryland 21702-1201 USA

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