|Item||Catalog #||Description||Quantity||Price (USD)|
|Bacterial Strain||13333||Bacteria in agar stab||1||$75|
This material is available to academics and nonprofits only.
Growth in Bacteria
Growth instructionsGrow at 32oC. Unable to grow at 42oC.
SpeciesThis is a strain, not a plasmid
Mutationgenotype: strA galK gyrB221 (CouR) gyrB203 (ts). CouR mutation is Arg136Cys. ts mutation is Pro171Ser.
- Cloning method Restriction Enzyme
- 5′ sequencing primer n/a (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
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).
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:N4177 strain was a gift from Martin Gellert (Addgene plasmid # 13333)
For your References section:Regulation of the genes for E. coli DNA gyrase: homeostatic control of DNA supercoiling. Menzel R, Gellert M. Cell. 1983 Aug . 34(1):105-13. 10.1016/0092-8674(83)90140-X PubMed 6309403