| Method |
The strains used throughout this
work were the isogenic prototrophic strains FB8 (relA +)
and FB8r (relA) (Uzan and Danchin 1978). Cells were
grown in a synthetic medium containing 0.1 mM phosphate
as already described (Danchin and Dondon 1980) and labeled
with 32P (150 microCi of carrier free, neutralized 32P.
phosphate, Amersham, UK) 90 min before addition of inhibitors
(Danchin and Dondon 1980). Chromatography of Phosphorylated Molecules: Small
phosphorylated molecules were submitted to chromatography
on PEI-cellulose sheets (Machery-Nagel, Diiren, FRG)
in either one or two dimensions. Sheets were then autoradiographed
and relevant spots were cut out and counted
for radioactivity in a liquid scintillation counter. |