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To evaluate whether ΔNp63 was required for stratified epithelial commitment, stable ES cell clones that express a small hairpin (sh) RNA inhibiting p63 expression were produced.
In vivo clonal analysis in mice and in vitro 3D modelling show that clones that express high levels of COL17A1, which divide symmetrically, outcompete and eliminate adjacent stressed clones that express low levels of COL17A1, which divide asymmetrically.
A cDNA clone library constructed with mRNA from SV40-transformed human fibroblasts and this vector (about 1.4 X 10 6) clones) yielded full-length cDNA clones that express hypoxanthine-guanine phosphoribosyltransferase (Jolly et al., Proc. Natl. Acad. Sci. U.S.A., in press).
By RT-PCR we identified clones that express wild type Tcf3 mRNA (Fig. 4D).
This might explain the selection of B cell clones that express antibodies reacting independent of the native antigen conformation.
To test the latter possibility, PLB-985 cell clones that express normal or diminished amounts of Hv1 were differentiated for 7 days in DMFA containing, low-serum medium.
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Ectopic expression clones that expressed salm, Delta, knirps, knirps and Notch-RNAi, or Notch Act were induced in 2-4 day old animals by heat shock at 37° for 10 minutes; wandering L3 larvae were dissected for analysis after 48 hours.
We generated small clones that expressed FLAG-Fbxl7, which enabled us to examine the borders between FLAG-Fbxl7-expressing cells and wild-type cells.
However, application of anti-repressor elements, which increase promoter activity, did induce the formation of clones that expressed proteins at high levels.
All six clones that expressed a cellulase in initial functional screening were tested for their thermal stability.
Based on the dot blot results, two clones that expressed a high level of the VRC01 antibody were selected from both the α and M strains for expression in shake flasks.
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