Part:BBa_K3570021:Experience

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Description

NLS-VP16-EL222 is a blue light-sensitive transcription factor. EL222 contains an LOV domain that forms homodimers in the presence of blue light and induces binding to a C120 consensus DNA motif(TAGCCTTTAGTCCATG). This part has been developed for use in the yeast S.cerevisiae and contains a nuclear localization sequence and VP16 activation factor functional in S.cerevisiae.

Usage

NLS-VP16-EL222 can be expressed in S.cerevisiae as a trans-regulatory factor for a promoter containing a C120 motif in the upstream activation position relative to the core promoter. In blue light, NLS-VP16-EL222 dimerizes and binds to the C120 motif, and a conjugated VP16 domain will promote transcription of downstream elements.

Characterization of expression from an episomal plasmid

NLS-VP16-EL222 was expressed constitutively using the native promoter ACT1p in S.cerevisiae strain BY4741 from an episomal plasmid alongside a minimal C120 containing promoter upstream of the fluorescent protein mKO(Figure 1). BY4741 containing this construct was exposed to blue light as well as darkness for 6 hours, and compared to BY4741 without the construct present

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