We have optimized the expression of several far-red fluorescent reporters in M. tuberculosis by combining codon-biased synthetic genes with optimal promoters and replicons to produce reporter strains that can be detected with great sensitivity in vitro.
In total eight embryos containing the 295C transgene showed robust staining of the ZPA in both forelimbs and hindlimbs, indicating that the construct produced reporter expression in the normal pattern of Shh (Fig. 5A).
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The promoter was first cloned into pCR®2.1-TOPO®, then released by EcoRI digestion and re-cloned into EcoRI-linearized pd2EGFPΔCMV producing the reporter plasmid pd2EGFP-Meq.
The performance of BYL compared to commercial WGEs was investigated by producing the reporter proteins eYFP and luciferase in both systems and measuring fluorescence, which is much faster than standard methods and does not involve radioactive labeling.
Previously reported 5′ deletions to positions −509 (pRJM106) and −280 (pRJM160) failed to produce any reporter gene expression [ 12].
The biosensing system takes advantage of the recognition of the regulatory protein, ArsR, for arsenite and antimonite to produce the reporter protein, which in this case is GFPuv.
To assess whether the in vivo-selected AAV-2 vectors mediate gene expression in tumor tissue in vivo, we produced luciferase reporter vectors displaying the selected peptide sequences.
The MultiNotch MS3 scan typically produced ∼2 500 reporter ions per MS3 spectrum.
Biolistic gene transfer to mouse skin produced peak reporter gene expression one day after transfer.
