Addgene - N4177 strain Plasmid Data

Bacterial strain 13333: N4177 strain

Gene/insert name:
None
Species:
This is a strain, not a plasmid
Mutation:
genotype: strA galK gyrB221 (CouR) gyrB203 (ts). CouR mutation is Arg136Cys. ts mutation is Pro171Ser.
Vector backbone:
n/a
(Search Vector Database)
5' sequencing primer:
n/a List of Sequencing Primers
Bacterial resistance(s)
coumermycin
Growth strain(s)
Unknown
Growth temperature (℃):
30
Growth instructions:
Grow at 32oC. Unable to grow at 42oC.
High or low copy:
Unknown
Principal Investigator:
Martin Gellert
Terms and Licenses:
MTA

Comments:

The strain was derived as follows: a culture of E. coli N99 galK strA was mutagenized with 1-methyl-3'-nitrosoguanidine and Cou-resistant cells were selected on LB agar containing 25ug/mL Cou and 10mM sodium pyrophosphate. These colonies were then screened for failure to grow on LB agar at 42oC. One such strain, N4175, was used as a donor for P1 transduction into N99. The transductant strain, N4177, is Cou-resistant at 32oC and is unable to grow at 42oC.

GyrB protein partly purified from N4177 cells grown at 32oC, when added to wild-type GyrA protein, yields a DNA gyrase complex whose supercoiling activity is Cou-resistant at 25oC and temperature sensitive (less than 1% of the activity of the normal enzyme at 42oC).

This plasmid has not been sequence verified. If you would like Addgene to sequence a portion of this plasmid before you request it, please contact us.

Article: Regulation of the genes for E. coli DNA gyrase: homeostatic control of DNA supercoiling. Menzel et al (Cell. 1983 Aug . 34(1):105-13. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 13333" in your Materials and Methods section.