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Agrobacterium tumefaciens strain LBA4404 was transformed with plasmid pGSA1131 containing chloramphenicol acetyltransferase gene (cat) for bacterial selection, a plant selectable marker phosphinothricin acetyl transferase gene (bar), and β- glucuronidase gene (gus).
Transformed bacteria were selected on LB-plates containing either ampicillin, or kanamycin depending on respective bacterial selection markers (Table 1).
This display system may complement currently available bacterial selection techniques.
Characterizing the selected variants of the semi-rational library demonstrated the potential of the innovative bacterial selection approach.
The original vector contains the spectinomycin/streptomycin adenylyltransferase (aadA) gene for bacterial selection and the hygromycin phosphotransferase gene (hpt) for transformed plant selection.
However, some bacterial strains used to transform plants, such as Agrobacterium rhizogenes strain K599, have elevated tolerance to spectinomycin and streptomycin, thus making bacterial selection of pGWB533 inefficient.
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Some of the consequences of the bacterial culture selection on capital and operating cost are also presented.
Rods in the process of cell division and spirillum-shaped bacteria are captured twice as well as single cells, indicating that bacterial size selection is due to physical capture by the filtering apparatus and not by surface properties of the prey (primary source).
However, complications related to protein-folding efficiencies of the gene insert in fusion with fluorescent protein reporters prevent effective on-plate bacterial colony selection leading to its limited use.
Bacterial plasmid selection and maintenance was performed using 100 mg/L kanamycin or 25 mg/L zeocin (Invitrogen).
This was presumably due to the increased lifespan of treated mice, allowing for additional rounds of bacterial replication, selection, and mutagenesis during antibiotic exposure.
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