Part:BBa_K2100061:Experience

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Applications of BBa_K2100061

The MIT iGEM team used pENTR hEF1a-Sv40 to stop transcription of a fluorescent protein following it. We used this part to build hEF1a-SV40:eYFP.

We transfected hEF1a-Sv40: eYFP into HEK293 cells along with a transfection marker, constitutively expressed red fluorescent protein mKate. Unfortunately, we saw high basal expression of the yellow fluorescent protein following the transcriptional stop signal in this entry vector.

Because hEF1a-SV40:eYFP expressed similar amounts of yellow fluorescence as the constitutively expressed mKate and eYFP control, this data shows that the transcriptional stop signal does not work as expected. Further testing can be done to determine if multiple copies of SV40 will decrease expression of the following gene.

User Reviews

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