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Revision as of 22:26, 14 October 2018
VERSION 1.0
余戎镇 正在添加底部内容
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Results
SynRT toolkits version 1.0 is the first generation thermosensors collection we provide to users. It contains 48 heat-induced RNA-based thermosensors.
We constructed in a total of 150 heat-inducible RNA thermosensors using goldengate assembly described in the Construction page. And the measurement method was describled in Measurement page.
We measured the activities of these thermosensors at six different temperatures: 29, 31, 35, 37, 39 and 42℃.Upon screening, we selected 48 heat-inducible RNA thermosensors out of the 150 members. And figure 1 shows the 48 thermosensors' activity. The positive control we designed won't form stem-loop structure with SD sequence by software prediction, and the experimental charaterization is described in Measurement page. The normalized fluorescence are used to describe the change of activity compared with positive control. We find that all of these thermosensors' expression level increase at elevated temperature, and the extent of increase relative to temperature is different.
To quantify different features of this library, we computed the fold-change of the response in the given temperature range. As the figure shows, from 29 to 37℃, the fold change ranges from x fold to x fold. And from 37 to 42℃, the fold change ranges from x fold to x fold. These fold-changes were higher than positive control. These results show that there is a diverse set of heat-inducible RNA thermosensors in toolkit.
Discussion: as these results show, the heat-inducible RNA thermosensor we designed can work. The fluorescence value increases with temperature elevated. Besides, the difference in fluorescence intensity and the rate of increase points to the diversity in thermosensor response. We think the sequence change in stem length, loop size, and mismatched or bulges in the stem give rise to the diversity in thermosensor response.
Combined with our human practice work we have mentioned in the Overview page. We also have built switching behavior fitting model to describe the continuous switching behavior of thermosenors.
>For more details about our model, you can visit Model page.< From the fitted curve, we can get the melting temperature, intensity and sensitivity of thermosensors. Based on these datas, we classified thermosensors, and develop a search engine -- SynRT Explorer. You can visit our Search Engine page to use it.
You can see the individual result as followed:
- BBa_K2541001
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- BBa_K2541003
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- BBa_K2541038
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