Journal: Scientific Reports
Article Title: Transcription Elongation Factor GreA Plays a Key Role in Cellular Invasion and Virulence of Francisella tularensis subsp. novicida
Figure Lengend Snippet: Analysis of the F. novicida greA locus. ( A ) Alignment of four Gram-negative bacterial GreA amino acid sequences using the MegAlign program of the DNAstar Lasergene package (version 10). Identical amino acid residues are shown in black. The cross-link with the RNA 3′-terminus is shown in a red box and the conserved acidic residues (D43 and E46) required for GreA activity are shown in green boxes. ( B ) Schematic illustration of the gene arrangement at the greA locus. ( C ) Cotranscription of the greA locus genes determined with RT–PCR. The pepA-–guaB (a), guaB– FTN_0662 (b), FTN_0662 –pgi (c), pgi–fimT (d), fimT–greA (e), and greA–uvrA (f) junctions were amplified using DNA, DNA-free RNA, or cDNA as the template. The images were acquired by the gel imaging system (LIUYI, Beijing, China). The experiment was repeated twice. The sizes of the molecular markers are indicated at the side in kbp.
Article Snippet: The Ribo-Zero™ Magnetic Kit (Epicentre, Madison, WI, USA) was used to remove the rRNA, and the rRNA-depleted RNA was used to generate cDNA libraries with the NEBNext Ultra™ Directional RNA Library Prep Kit for Illumina® (NEB).
Techniques: Activity Assay, Reverse Transcription Polymerase Chain Reaction, Amplification, Imaging