Measuring Plasmid

We recloned pSB1A10 to improve its features as a measuring plasmid to evaluate terminators in vivo using fluorescent proteins as reporters. RFP which was known to contain an RNase restriction site was exchanged against mCherry which combines good expression yield, short maturation times and an acceptable and well-characterized quantum yield. For easy introduction of the terminator to be evaluated, we ?? ?? ??

Aptamer

We also add an malachitegreen-binding aptamer to the partsregistry which allows evalutation of terminators using in vitro transcription. Upon binding of malachitegreen to the aptamer the fluorescence increases 2360-fold leading to an significant increase over the whole transcription time. With an emission maximum at 652 nm, aptamer-bound malachitegreen fluoresces at longer wavelength than most dyes and does not interfere with those.