genopar.org), and sub-cloned into the vector pLAFR3.18 digested with EcoRI-HindIII to yield plasmid pACB210. Construction of chromosomal amtB::lacZ transcriptional fusions To construct amtB – lacZ transcriptional fusions, the suicide plasmid pSUPamtBClacZ was introduced by conjugation, using E. coli strain S17.1 as the donor, Selleckchem Inhibitor Library into H. seropedicae strains SmR1, LNglnKdel and LNglnB resulting in the strains LNamtBlacZ, LNglnKamtBlacZ
and LNglnBamtBlacZ, respectively. Genomic DNA hybridization confirmed the presence of the cassette lacZ- KmR in the amtB gene (data not shown). Acknowledgements We are grateful to the GENOPAR consortium for providing plasmids, and to Roseli Prado, Julieta Pie and Valter Belnacasan price Baura for technical assistance. We are also grateful
to Dr. Geoffrey Yates for reading the manuscript. This work was supported by INCT-FBN/CNPq/MCT, Institutos do Milênio, PRONEX, CAPES, CNPq and Fundação Araucária. Electronic supplementary material Additional file 1: Immunoblot analysis of H. seropedicae PII proteins. (DOC 79 KB) References 1. Arcondeguy T, Jack R, Merrick M: P(II) signal transduction proteins, pivotal players in microbial nitrogen control. Microbiol Mol Biol Rev 2001, 65 (1) : 80–105.PubMedCrossRef 2. Forchhammer K: P-II signal transducers: novel functional and structural insights. Trends Microbiol 2008, 16 (2) : 65–72.PubMed 3. Jiang P, Ninfa AJ: Escherichia coli PII signal transduction protein controlling nitrogen assimilation acts as a sensor of adenylate energy charge in vitro. Biochemistry 2007, 46 (45) : 12979–12996.PubMedCrossRef 4. He LH, Soupene E, Ninfa A, Kustu S: Physiological role for the GlnK protein of enteric bacteria: Relief of NifL inhibition under nitrogen-limiting conditions. J Bacteriol 1998, 180 (24) : 6661–6667.PubMed 5. Jack R, De Zamaroczy M, Merrick M: The signal transduction protein GlnK is required for NifL-dependent nitrogen control of nif gene expression
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