Fig. 5. RT-PCR analysis and dot-blot hybridization
of the pegD (a) and pegA (b) in S.
macrogoltabida strain 203. M, lEcoT14I
marker; +, positive control using total DNA
from strain 203 as a template; ”, negative
control prepared by skipping the reversetranscription
step; G and P, RNA samples
were isolated from glucose- and PEG 4000-
grown cells, respectively. The conditions
used for pre-hybridization, hybridization,
washing and detection followed the instructions
recommended by the manufacturer.
(c) The quality of isolated RNA samples was
checked by electrophoresis. Ten micrograms
of isolated total RNA sample was loaded in
a 0.8% agarose gel and electrophoresed in
20 mM MOPS buffer (pH 7.0) containing
1 mM EDTA, 5 mM sodium acetate and 4%
formaldehyde. The gels were stained with
10 mg ethidium bromide ml”1.
344 Microbiology