Journal: Microorganisms

Article Title: Identification of Phage RNA Polymerases That Minimize Double-Stranded RNA By-Product Formation and Their Characterization via In Vitro Transcription

doi: 10.3390/microorganisms14030564

Figure Lengend Snippet: Expression analysis of 44 RNA polymerase (RNAP) variants. All RNAP variants were expressed in E. coli BL21(DE3), and crude lysate samples were analyzed via SDS-PAGE. An empty vector control (IPTG-induced cells harboring an empty pET28a vector) was used for the gels to determine host-derived background protein bands. The gel images show sections of gels at sizes relevant for the respective protein. The samples were collected after four hours of induction with 0.5 mM IPTG. The size of all proteins is given in kilodaltons (kDa) and indicated below each SDS gel.

Article Snippet: BL21(DE3) Competent E. coli , C2527H , NEB Inc..

Techniques: Expressing, SDS Page, Plasmid Preparation, Control, Derivative Assay, SDS-Gel

Journal: The Journal of Biological Chemistry

Article Title: Biochemical characterization of the first step in sulfonolipid biosynthesis in Alistipes finegoldii

doi: 10.1016/j.jbc.2022.102195

Figure Lengend Snippet: Identification and purification of cysteate acyl-ACP transferase. A , TLC separation of 3-keto[ 14 C]capnine (3[ 14 C]KC) arising from SulA activity using cellular lysates from Escherichia coli cells expressing nothing (empty vector control), alfi_1224 (SulA), or alfi_1465 . The thin-layer plate is representative of four independent experiments. B , purified SulA migrates as a single 85 kDa species ( V e = 76.6 ml) based on a calibrated HiLoad 16/600 Superdex 200 column (98.1 kDa theoretical homodimer mass). Left inset , molecular weight calculation using calibration standards described under . Right inset , SDS gel electrophoresis shows SulA purity (∼48 kDa) along with molecular weight standards. C , the sedimentation velocity profile (fringe displacement) was fit to a continuous sedimentation coefficient distribution model c(S). SulA sedimented as a 98.9 kDa homodimer with a sedimentation coefficient of 5.11 S. ACP, acyl carrier protein; SulA, cysteate acyl-ACP transferase.

Article Snippet: E. coli strain BL21(DE3) cells harboring the pET-SulA or pET-SulA(K281A) expression plasmids were grown in 2 l LB medium with 50 μg/ml kanamycin (Gold Biotechnology) at 37 °C and 200 rpm shaking to A 600 = 0.6.

Techniques: Purification, Activity Assay, Expressing, Plasmid Preparation, Molecular Weight, SDS-Gel, Electrophoresis, Sedimentation

Journal: Nature chemical biology

Article Title: Monitoring methionine sulfoxide with stereospecific mechanism-based fluorescent sensors

doi: 10.1038/nchembio.1787

Figure Lengend Snippet: E. coli cells expressing MetSOx ( a ) or MetROx ( b ) were incubated with MetO for ~30 min, and then rinsed to remove the oxidant. Arrows indicate the addition (↓) of MetO and rinsing (↑) the cells for two representative experiments (Assay 1 and Assay 2). MetSOx ( c ) and MetROx ( d ) oxidized fractions calculated from (n = 3), using and , respectively, described in the “ online methods ” section.

Article Snippet: SoluBL21 TM E. coli (Gelantis, San Diego, CA) cells were transformed with an indicated expression vector and grown in Luria-Bertani containing ampicillin (50 μg.ml −1 ) at 37°C.

Techniques: Expressing, Incubation