Part:BBa_K2100072:Experience

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Applications of BBa_K2100072

The MIT iGEM team used pEXPR hEF1a-Sv40: eYFP to show that SV40 would stop transcription of constitutively expressed eYFP following it.

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 in this expression 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 yellow fluorescent protein following it.

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