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Previously, the BFPvvD7 mutant generated by directed evolution displayed a fourfold enhancement in fluorescent intensity.
Consider again the FM of Fig. 4. Figure 5 contains an example of mutant generated by the operator AFS (Add Feature to a Set relation (solitary feature to grouped)).
Further molecular analysis of a mutant generated by imprecise excision of the P element reveals that a partial deletion of the PTE and neighboring upstream genomic sequences results in a similar loss of enhancer-directed Abd-B expression.
This plasmid was introduced in BAB-parental and the deletion mutant generated by allelic exchange was selected by Nal and sucrose resistance and Gm sensitivity and by PCR using oligonucleotides olsA-F1 and olsA-R4 which amplify a fragment of 870 bp in the deletion strain and a fragment of 1464 bp in the parental strain.
With a kinase-defective (KD) BCR-ABL mutant, generated by substitution of the critical Lysine in the ATP-binding site, nuclear accumulation is achieved by treatment with LMB alone, suggesting that the NLS is active in BCR-ABLKD [22] (supplementary Figure S1).
As a control, a catalytically inactive mutant generated by a His (599) to Ala mutation was included.
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cbpA and cbpB mutants, generated by gene disruption, showed normal growth, development and spore germination.
More recently, this approach has been used to interrogate a collection of conditional mutants generated by random mutagenesis to identify canonical splicing mutants.
Allelic exchange mutants generated by double cross-over homologous recombination were selected by two-day microaerobic incubation on BHI agar plates containing 500 μg/ml kanamycin.
To avoid positional effects of the transgene, exactly the same line was used in the wild type and mutants, generated by crossing.
In vegetative tissues, SlIAA9-crispr exhibited the leaf phenotype typical of other SlIAA9 mutants generated by chemical, physical, or RNAi induced mutation8.
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