molecular weight marker generuler tm 1 kb plus dna ladder  (Thermo Fisher)

Journal: Scientific Reports

Article Title: Unraveling the role of the CbrA histidine kinase in the signal transduction of the CbrAB two-component system in Pseudomonas putida

doi: 10.1038/s41598-019-45554-9

Figure Lengend Snippet: Genomic organisation of the cbrX-cbrA-cbrB cluster in P. putida . ( A ) Graphic representation of the gene organisation of cbrX ( PP5704 in red), cbrA ( PP4695 in purple), cbrB ( PP4696 in yellow) and crcZ ( PPmr53 in white) in Pseudomonas putida KT2442. The promoters for cbrA , cbrB and crcZ are depicted as black arrows. ( B ) DNA sequences for the promoter region and of cbrX and 5′ of cbrA in Pseudomonas putida KT2440. Underlined sequences correspond to the putative -10 and -35 boxes, a predicted Crc binding site is shown in italics and with a dotted underline, and the putative RBS for cbrX “CGAGG” is boxed. The coding sequence for cbrX is represented in red characters, that of cbrA is shaded in purple, and the overlapping sequences for cbrX and cbrA are shaded in grey. The start and stop codons are highlighted in bold. ( C ) cbrX and cbrA are co-transcribed in the same transcriptional unit. RT-PCR of 25, 5 and 1 ng of cDNA from a culture KT2442 growing in OAA was amplified with different combination of the primers represented in panel A; primers 1 (RT-PcbrXA_fwd2) and 2 (RT-cbrX_rev) (lines 1 to 3), primers 1 (RT-PcbrXA_fwd2) and 4 (RT-CbrA_rev) (lines 4 to 6) and primers 3 (RT-CbrA_fwd) and 4 (RT-CbrA_rev) (lines 7 to 9). Positive and negative PCR controls (+and −, respectively) were performed with genomic or no DNA for each pair of oligos (see Methods). M is the DNA molecular weight marker GeneRuler TM 1 Kb Plus DNA ladder (ThermoFisher Scientific).

Article Snippet: M is the DNA molecular weight marker GeneRuler TM 1 Kb Plus DNA ladder (ThermoFisher Scientific). cbrX encodes a small 58 amino acids peptide of unknown function and is well conserved within the genera Pseudomonas and Azotobacter from the Pseudomondaceae family (Fig. ).

Techniques: Binding Assay, Sequencing, Reverse Transcription Polymerase Chain Reaction, Amplification, Molecular Weight, Marker