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In previous studies, transgene expression was analysed in clonal plant replicates generated long time after the integration event [ 10, 17].
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We first studied transgene expression by analyzing the pattern of EGFP expression in Eno2-ΔGR/EGFP mice maintained in doxycyline-free condition from birth to adulthood.
In the present study, transgene expression remained high after 60 days and was still detectable after 5 months (data not shown).
We previously created a number of RMCE integration sites termed RL1 to RL6 that can be used to study transgene expression in mouse erythroleukemia (MEL) cells.
MFI was analyzed by flow cytometry to study transgene expression levels induced by the CMV and CAG promoters in DF-1 cells.
For studies of transgene expression in genetic backgrounds lacking endogenous VAP, VAPΔ166 /FM7Kr-GFP; elav-GAL4 females were crossed to w/Y; UAS-VAP males (either wild type, P58S, or the recombined chromosome containing both transgenes).
In similar studies exploring transgene expression in skin after non-electroporative intradermal injection, expression was detectable with an in vivo imaging system [26] 10 hrs after DNA administration, and after 6 hrs using the more sensitive in vitro detection assay [15].
The cloning procedure can be used not only for efficient reduction of expression heterogeneity of such transgenes, but also as a useful tool for studies of transgene expression and other purposes.
The study of transgene expression is of vital importance whenever transgenic plants are produced.
In one study, Bcl2 transgene expression did not block superantigen negative selection (Sentman et al., 1991) while another group clearly saw a rescue (Siegel et al., 1992).
Although it is difficult to select the homozygous line in the progeny derived from a transformant with high copy number for the purposes of genetic study, exogenous transgene expression level is still relatively stable in the next generations as we observed (Additional file 1: Figure S1).
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