Difference between revisions of "Part:BBa K4164020"
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The improvement of NAU-CHINA this year is to integrate TEV protease and Ssr tag to construct BBa_K4164020 in order to control the degradation of mRFP and resolve the leakage problem of BBa_J04450. | The improvement of NAU-CHINA this year is to integrate TEV protease and Ssr tag to construct BBa_K4164020 in order to control the degradation of mRFP and resolve the leakage problem of BBa_J04450. | ||
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TEV cleavage site (ENLYFQS) and ssr tag (AANDENYAAV) are inserted into the C-terminus of RFP in turn (Fig.1). Meanwhile, the TEV protease is expressed also driven by Plac. The ssr tag can lead to the degradation of RFP expressed without IPTG. When induced by IPTG, the TEV protease can be inductively expressed, and cleave specifically at the cleavage site, which results in the loss of ssr tag in RFP, and RFP lost the degradation tag will no longer be degraded and begin to work. | TEV cleavage site (ENLYFQS) and ssr tag (AANDENYAAV) are inserted into the C-terminus of RFP in turn (Fig.1). Meanwhile, the TEV protease is expressed also driven by Plac. The ssr tag can lead to the degradation of RFP expressed without IPTG. When induced by IPTG, the TEV protease can be inductively expressed, and cleave specifically at the cleavage site, which results in the loss of ssr tag in RFP, and RFP lost the degradation tag will no longer be degraded and begin to work. | ||
Revision as of 12:50, 8 October 2022
Improvement
The improvement of NAU-CHINA this year is to integrate TEV protease and Ssr tag to construct BBa_K4164020 in order to control the degradation of mRFP and resolve the leakage problem of BBa_J04450.
TEV cleavage site (ENLYFQS) and ssr tag (AANDENYAAV) are inserted into the C-terminus of RFP in turn (Fig.1). Meanwhile, the TEV protease is expressed also driven by Plac. The ssr tag can lead to the degradation of RFP expressed without IPTG. When induced by IPTG, the TEV protease can be inductively expressed, and cleave specifically at the cleavage site, which results in the loss of ssr tag in RFP, and RFP lost the degradation tag will no longer be degraded and begin to work.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1694
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 814
Illegal AgeI site found at 926 - 1000COMPATIBLE WITH RFC[1000]