Difference between revisions of "Part:BBa K3317067"
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RNA thermometer, by subjecting the cells to temperatures greater than 37 ℃ the expression of the proteins placed downstream is allowed, this sequence is an improvement of (<partinfo>K1140006</partinfo>) | RNA thermometer, by subjecting the cells to temperatures greater than 37 ℃ the expression of the proteins placed downstream is allowed, this sequence is an improvement of (<partinfo>K1140006</partinfo>) | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | < | + | The primary objective was to perform an <i>in silico</i> design to reduce the range of operation of previously reported thermometers (28-37° C), obtaining a more precise range and allowing greater control of the system. |
| − | <span class='h3bb'>Sequence and Features</span> | + | |
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| + | Tests were performed in conjunction with the (<partinfo>K1140006</partinfo>) thermometer from 25 to 40° C, measuring at 586/610 nm (excitation/emission for mCherry) and 660 nm to obtain optical density of the culture and normalize results, which showed that the new thermometer controlled the expression of the reporter in a more precise range, like the predicted theoretically. The <i>in silico</i> design allows the optimization of regulatory elements, such as RNA thermometers, reducing the number of experiments to a minimum. | ||
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| + | ==<span class='h3bb'>Sequence and Features</span>== | ||
<partinfo>BBa_K3317067 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3317067 SequenceAndFeatures</partinfo> | ||
| + | <partinfo>BBa_K1140006</partinfo> was redesigned in the area of the U6 thermometer, in order to reduce the operation temperature (28-37° C). The new piece, <partinfo>BBa_K3317067</partinfo>, showed as RNA Thermometer Pro in the features, consists of 121 nucleotides, with a Hairpin formation in the position 91 to 101, before the RBS (AGGAGA), allowing greater control of the system. | ||
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K3317067 parameters</partinfo> | <partinfo>BBa_K3317067 parameters</partinfo> | ||
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Latest revision as of 22:52, 21 October 2019
37 C inducible RNA thermometer
RNA thermometer, by subjecting the cells to temperatures greater than 37 ℃ the expression of the proteins placed downstream is allowed, this sequence is an improvement of (BBa_K1140006)
Usage and Biology
The primary objective was to perform an in silico design to reduce the range of operation of previously reported thermometers (28-37° C), obtaining a more precise range and allowing greater control of the system.
Tests were performed in conjunction with the (BBa_K1140006) thermometer from 25 to 40° C, measuring at 586/610 nm (excitation/emission for mCherry) and 660 nm to obtain optical density of the culture and normalize results, which showed that the new thermometer controlled the expression of the reporter in a more precise range, like the predicted theoretically. The in silico design allows the optimization of regulatory elements, such as RNA thermometers, reducing the number of experiments to a minimum.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 63
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
BBa_K1140006 was redesigned in the area of the U6 thermometer, in order to reduce the operation temperature (28-37° C). The new piece, BBa_K3317067, showed as RNA Thermometer Pro in the features, consists of 121 nucleotides, with a Hairpin formation in the position 91 to 101, before the RBS (AGGAGA), allowing greater control of the system.