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Table 1 The strains and plasmids utilized in this work

From: Tools for genetic manipulation of the plant growth-promoting bacterium Azospirillum amazonense

Bacterial strains Genotype Reference  
Escherichia coli XL1-Blue recA1, endA1, gyrA96, thi-1, hsdR17(rK-, mK+), supE44, relA1, lac, [F', proAB, lacIqZΔM15::Tn10(tetr)] Stratagene  
Azospirillum amazonense Y2 wild type EMBRAPA-RJ  
Azospirillum amazonense delK Y2 derivative, ΔglnK This work  
Plasmids Relevant characteristics Reference Purpose
pUC18 cloning vector, ampr, lacZα Clontech cloning procedures
pAAGLNK pUC18 derivative containing the SalI genomic fragment of the glnK region This work glnK isolation
pGLNBA pUC18 derivative containing the amplification product generated from the revsf_glmBint and gln_AA_do primers This work glnB-glnA intergenic region and partial glnA isolation
pRK2013 ColE1 ori, kanr, mob, tra [45] helper plasmid for conjugation experiments
pHRGFPGUS pBBR1 ori, kanr, mob, gfpmut3 [46] DNA transfer evaluation, reporter vectors construction
pPZP201BK pVS1 ori, ColE1 ori, kanr, mob [47] construction of pPZPLACEYFP
pEYFP pUC ori, eyfp, ampr Clontech contruction of reporter plasmids
pPZPLACEYFP PvuII-EcoRI fragment from pEYFP (containing the lac promoter-eyfp gene fusion) cloned into the EcoRV-EcoRI sites of pPZP201BK This work DNA transfer evaluation, positive control in the reporter assay
pK19MOBSACB Integration vector, kanr, ColE1 replication origin, mob, sacB, lacZα [48] glnK mutagenesis
pKΔK pK19MOBSACB derivative containing the flanking regions of the glnK gene joined by Crossover PCR This work glnK mutagenesis
pAATEYFP pEYFP derivative containing the BglII-NcoI fragment of pAAGLNK cloned into its NcoI-BamHI sites This work construction of pHRAATEYFP
pPBEYFP pEYFP derivative containing the intergenic region between the bcr protein superfamily gene and the glnB gene cloned into its NcoI-BamHI sites This work construction of pHRPBEYFP
pPKEYFP pEYFP derivative containing the intergenic region between the ubiH and the glnK genes cloned into its NcoI-BamHI sites This work construction of pHRPKEYFP
pHRAATEYFP HindIII-EcoRI fragment from pAATEYFP (containing the aat promoter-eyfp gene fusion) joined with the 5.8 kb HindIII-EcoRI fragment of pHRGFPGUS This work promoter evaluation
pHRLACEYFP PvuII-EcoRI fragment from pEYFP (containing the lac promoter-eyfp gene fusion) joined with the 7.4 kb EcoRV-EcoRI fragment of pHRGFPGUS This work promoter evaluation
pHRPBEYFP HindIII-EcoRI fragment from pPBEYFP (containing the glnB promoter-eyfp gene fusion) joined with the 5.8 kb HindIII-EcoRI fragment of pHRGFPGUS This work promoter evaluation
pHRPKEYFP HindIII-EcoRI fragment from pPKEYFP (containing the glnK promoter-eyfp gene fusion) joined with the 5.8 kb HindIII-EcoRI fragment of pHRGFPGUS This work promoter evaluation
pHREYFP HindIII-EcoRI fragment from pEYFP (containing the eyfp gene) joined with the 5.8 kb HindIII-EcoRI fragment of pHRGFPGUS This work promoter evaluation