duncant at cross.bmb.hscsyr.edu
Mon Jan 6 19:15:26 EST 1997
Pascal Mertens wrote:
> f1 is the same bacteriophage that fd or M13 (discovered at the same time
> in 3 different labs).
> Infect E.c. cells grown in terrific broth (OD= 0.125 to 0.25 for a
> dilution 10 of the culture), the cells beeing left 5 min on table to
> make the pilus regenerated. Then mix equal volumes (10 microL) of
> E. coli cells and phage dilutions ; incubated 10 min on table; add 1
> ml of 2xYT medium and incubate 20 min at 37 deg. Plate. That's it.
> Hope this helps
Of course, the expression of F-pili on the surface of E. coli cells
requires genes expressed from an F' episome, so you should be sure the
strain you use harbors an F'. Most F'-containing strains allow a growth
selection for the presence of the F', and many "crippled" F' (with traD
mutation) can be lost easily if not selected for. Also, expression of
F-pili is sensitive to growth temperature (I think it's best between
35-40 deg. C).
Thomas M. Duncan
Dept. Biochemistry & Molecular Biology
SUNY Health Science Center
750 E Adams St, Syracuse, NY 13210
Email: duncant at vax.cs.hscsyr.edu or duncant at cross.bmb.hscsyr.edu
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