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Apart from a higher plating density, the use of multiple reporter genes has many additional advantages.
Our preliminary analysis of the multiple reporter genes substantiates that their expression appears to accurately reflect their endogenous genes expression, respectively.
To minimize false positives caused by self-activation, we used multiple reporter genes driven by different GAL4-responsive promoters and carefully determined the level of self-activation of each bait clone (see Materials and methods).
This DNA construct could then be used for mouse transgenesis to create an animal model where the multiple reporter genes would insert into a single locus, thus simplifying breeding schemes, yet expanding the capability and usage of the animal model.
Next, in order to investigate the in vivo immunological survival of BMSC genetically-modified with multiple reporter genes in immunocompetent mice, 2 × 105 BMSC-Luc/eGFP/Pac were implanted in the CNS of syngeneic ROSA26-L-S-L-luciferase transgenic mice (n = 8).
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However, TCRα LCR-driven transgenes display ectopic transcription in B cells in multiple reporter gene systems.
The main components of the ceh-63 expression pattern, as revealed by multiple reporter gene fusion arrangements, are DVC and uterus.
However, our preliminary observations of these transgenic lines suggest the outcome of this technical approach to design multiple reporter gene constructs has met with mixed success.
This problem has made us reconsider the design of transgenic reporter gene mice and appreciate the great value in establishing a methodology that would result in the generation of a single DNA fragment containing multiple reporter gene elements.
Spurdle et al (2002) evaluated multiple reporter gene constructs, but found no differences in the transcription between the putative disease and wild-type allele; the results of other functional studies have been equivocal (Ball et al, 1999; Westlind et al, 1999; Amirimani et al, 2003; Rodriguez-Antona et al, 2005).
Finally, a panel of 15 bioluminescent E. coli containing multiple bacterial reporter genes associated with oxidative stress, DNA damage, heat shock, and efflux of excess metals was arrayed to screen a library of 420 pharmaceuticals [ 69].
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