N4177 strain
(Bacterial strain #13333)

• Depositing Lab: Martin Gellert
• Publication: Menzel et al Cell. 1983 Aug . 34(1):105-13.

Backbone

• Vector backbone: n/a

Growth in Bacteria

• Bacterial Resistance(s): Coumermycin
• Growth Temperature: 30°C
• Growth Strain(s): Unknown
• Growth instructions: Grow at 32oC. Unable to grow at 42oC.
• Copy number: Unknown

Gene/Insert

• 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.

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: n/a

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor 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).