Affiliation:
1. Laboratory for Nitrogen Fixation Research, Oregon State University, Corvallis, Oregon 97331
Abstract
A method has been developed for the rapid screening of
Rhizobium japonicum
colonies for hydrogenase activity based on their ability to reduce methylene blue in the presence of respiratory inhibitors and hydrogen. Hydrogen uptake-positive (Hup
+
) colonies derepressed for hydrogenase activity were visualized by their localized decolorization of filter paper disks impregnated with the dye. Appropriate responses were seen with a number of Hup
+
and Hup
−
wild-type strains of
R. japonicum
as well as Hup
−
mutants. Its specificity was further confirmed in selected strains on the basis of comparisons with chemolithotrophic growth and the presence of other genetic markers. Utilization of the method in identifying Hup
+
colonies among 16,000 merodiploid derivatives of the Hup
−
mutant strain PJ17nal containing cloned DNA fragments of the Hup
+
strain 122 DES has demonstrated its applicability as a screening procedure in the genetic analysis of the
R. japonicum
hydrogen uptake system.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
30 articles.
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