Difference between revisions of "Part:BBa K3570021:Experience"
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| + | Figure 2 demonstrates that in the presence of blue light, mKO expression is increased roughly 3 fold compared to darkness. mKO expression was also measured overtime for 6 hours(Figure 3) and a comparison was made for 100% blue light, 100% darkness, and 50% blue light-duty cycles. While constitutive blue light increased expression over time, and constitutive darkness decreased expression over time, 50% blue light maintained roughly constant expression, demonstrating the ability of this part to modulate dose-dependent expression. | ||
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===User Reviews=== | ===User Reviews=== | ||
Revision as of 03:48, 18 October 2021
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Applications of BBa_K3570021
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 (Figure 2)
Figure 2 demonstrates that in the presence of blue light, mKO expression is increased roughly 3 fold compared to darkness. mKO expression was also measured overtime for 6 hours(Figure 3) and a comparison was made for 100% blue light, 100% darkness, and 50% blue light-duty cycles. While constitutive blue light increased expression over time, and constitutive darkness decreased expression over time, 50% blue light maintained roughly constant expression, demonstrating the ability of this part to modulate dose-dependent expression.
User Reviews
UNIQe6afc89e9b1355f6-partinfo-00000000-QINU UNIQe6afc89e9b1355f6-partinfo-00000001-QINU