Difference between revisions of "Part:BBa K3423005"
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Protein anti-CRISPR-associated 1 (Aca1) is constitutively expressed by T7 promoter, Aca1 will subsequently inhibit the anti-CRISPR promoter, so the gene expression downstream anti-CRISPR promoter will be inhibited until Navitoclax (a compound that can inhibit Aca1 found by SUSTech_Shenzhen) is added artificially. Without the inhibition from Aca1, the expression of genes downstream anti-CRISPR promoter will increase significantly. Therefore, protein expression can be regulated artificially. | Protein anti-CRISPR-associated 1 (Aca1) is constitutively expressed by T7 promoter, Aca1 will subsequently inhibit the anti-CRISPR promoter, so the gene expression downstream anti-CRISPR promoter will be inhibited until Navitoclax (a compound that can inhibit Aca1 found by SUSTech_Shenzhen) is added artificially. Without the inhibition from Aca1, the expression of genes downstream anti-CRISPR promoter will increase significantly. Therefore, protein expression can be regulated artificially. | ||
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| + | We tested this composite part using green fluorescent protein as a reporter, the green fluorescence intensity is measured with a microplate reader:https://static.igem.org/mediawiki/parts/3/33/SUSTech_ShenzhenGFP.png | ||
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| + | When Aca1 is mutated at its 44th amino acid (Arginine to alanine, Aca1R44A),the difference expression level of green fluorescent protein could be easily observed with naked eyes:https://static.igem.org/mediawiki/parts/8/8a/SUSTech_ShenzhenR44A.png | ||
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Revision as of 11:24, 23 October 2020
a controlled protein expression system
Protein anti-CRISPR-associated 1 (Aca1) is constitutively expressed by T7 promoter, Aca1 will subsequently inhibit the anti-CRISPR promoter, so the gene expression downstream anti-CRISPR promoter will be inhibited until Navitoclax (a compound that can inhibit Aca1 found by SUSTech_Shenzhen) is added artificially. Without the inhibition from Aca1, the expression of genes downstream anti-CRISPR promoter will increase significantly. Therefore, protein expression can be regulated artificially.
We tested this composite part using green fluorescent protein as a reporter, the green fluorescence intensity is measured with a microplate reader:
When Aca1 is mutated at its 44th amino acid (Arginine to alanine, Aca1R44A),the difference expression level of green fluorescent protein could be easily observed with naked eyes:
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1057