By Jeffrey H. Miller
This quantity collates in a single resource technique for in vivo genetic engineering and for genetic research in quite a lot of micro organism. not just is Escherichia coli good lined, yet so are different rising bacterial structures
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Additional resources for Bacterial Genetic Systems
The advantages of the in vivo approach are (1) in some cases spontaneous mutants can be selected and (2), when not, the range of specificities of mutagenic agents and mutator genes allows the isolation of mutants with a wide variety of base alterations. Selecting or Screening for Mutants in Diploids Selecting for mutants in diploids was first used to obtain mutations in the/3 and/3' subunits of RNA polymerase. 27 Mutations of E. coli resistant to the antibiotic rifamycin occur in the gene for the/3 subunit, rpoB.
The agar is allowed to solidify, and the plate is incubated overnight at 37° in an upright position. The next day a single plaque ( - 10 6 phage) is picked with a microcapillary tube and mixed with 50/zl of an overnight bacterial culture. The mixture is incubated for 5 min at 37° to permit phage adsorption and then plated as described above onto an LCA plate that is prewarmed to 38°. The plate is immediately placed at 38° to prevent complete solidification of the top agar and is incubated for 5-6 hr at that temperature.
Schell, W. Symonds, and K. A. Stacey, Genet. Res. 13, 227 (1963). 26 Escherichia coli AND Salmonella typhimurium  ylase (dam) g e n e , 7'1° and mutations that permit PI d a m - mutants to grow better in a d a m - host (rev6 and rev7). The dam::Kan strain contains an insertion of the Tn903 kan r gene in dam, and AMB contains a 320-bp deletion in dam. Bacterial Strains P1 lysates can be made on most strains ofE. coli K12 that are recA + although the phage yield (25-150 infectious phage per cell) may vary significantly.