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We then compared the three multiple epitope presentations in a microarray assay using sera samples from milk allergic patients, confirming that a multiple presentation, in particular that of the tandem construct, allows for a more efficient characterization of IgE-binding fingerprints at a statistically significant level.
The tandem construct provides a sensitive measure of transcription termination in human cells.
Since the ribozymes contribute to the effectiveness of the tandem construct, it was necessary to confirm ribozyme cleavage in the cell.
Cell lines with a single integrated tandem construct showed strong expression of both FLUC and hRLUC reporters upon induction (compare minus and plus induction in Figure 1B).
PCR amplification with the same primers on a construct similar to the tandem construct but lacking ribozymes (TAN NR) yielded a PCR product (Figure 1C).
The FRET efficiency of a tandem construct can be calculated from the decreased donor lifetime (τϕ or τΜ) relative to the fluorescence lifetime of an unquenched donor according to equation 3.
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Circular permutated yellow fluorescent protein variants revealed that in the tandem constructs the orientation of the transition dipole moment influences the FRET efficiency.
To prevent generating such competitive transgenic progeny, we previously used tobacco (Nicotiana tabacum L). as a model for validating a transgenic mitigation (TM) mechanism using tandem constructs where a gene of choice is linked to mitigating genes that are positive or neutral to the crop, but deleterious to a recipient when in competition with the wild type.
Temperature profiles of Cel5A in mixture and alone were highly comparable proving that faster inactivation of tandem constructs is due to artificial fusion.
The deletion became necessary to obtain reliable expression of the tandem constructs.
Mutants and tandem constructs were generated using PCR with Pfu DNA polymerase by standard protocols.
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