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By R. Fani, M. Bazzicalupo, E. Gallori, L. Turbanti, M. Polsinelli (auth.), Prof. Dr. Walter Klingmüller (eds.)

ISBN-10: 3642707912

ISBN-13: 9783642707919

ISBN-10: 3642707939

ISBN-13: 9783642707933

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Additional resources for Azospirillum III: Genetics · Physiology · Ecology Proceedings of the Third Bayreuth Azospirillum Workshop

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Considering the Testriction sites mapped in the present study, there are no similarities between the restriction maps of Ali nifHDK and the restriction ma ps of nifHDK of A. brasilense or ~ pneumoniae. Complementation of K. pneumoniae nif mutants The cloned nif structural genes were tested for complementation of K. pneumoniae nifH, nifD and nifK mutants. The nifHDK genes of ~ lipoferum cloned in pFS393 and pFS13 were introduced into K. pneumoniae wild type and nif- mutants. pFS393 carries the 8 kb Pst1 fragment of plasmid pFS191.

Ml Te. Table 3 Nitrogenase activity of A. brasilense 7029 containing various pVK100 derivatives. 2 Nitrogenase activity was assayed under derepression conditions. 5%) 46 Strain 7029 has a Gln- Nif- phenotype and we measured the nitrogenase activity in the different transconjugants (Table 3). Cosmid pVK100 did not modify the nitrogenase activity of strains 7000 and 7029 (data not shown). Plamids pAB44 , -221 and -223 that restored glutamine independent growth restored a normal control of nitrogen fixation in 7029.

Preparation of DNA: Lysis of Azsopirillum strains and purification of chromosomal ONA was don e essentially as described by Marmur (17). 5%) and proteinase K (50 ~g/ml). ONA was further purified by cesiumchloride-ethidiumbromide (CsCI-Ethbr) equilibrium density-gradient centrifugation. Plasmid ONA was isolated as described by Rodriguez and Tait (18). Large quantities of plasmid ONA were obtained by the eleared lysate method followed by a final purification in a CsCI-Ethbr equilibrium density-gradient centrigugation (19).

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Azospirillum III: Genetics · Physiology · Ecology Proceedings of the Third Bayreuth Azospirillum Workshop by R. Fani, M. Bazzicalupo, E. Gallori, L. Turbanti, M. Polsinelli (auth.), Prof. Dr. Walter Klingmüller (eds.)


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