Ara-Inducible Cyan7427_1390g3 Expression System
A GAF domain of the cyanobacteriochrome (CBCR) Cyan7427_1390 with an intein/chitin-binding domain fused to the C-terminus for purification. Expression regulated by a pBAD promoter. As with most CBCRs, Cyan7427 requires a phycocyanobilin co-factor, and must therefore be expressed in a system with heme oxygenase (BBa_I15008) and PcyA (BBa_I15009). Cyan7427_1390g3 expresses well in E. coli and cells appear vibrant blue-green under normal light.
The ara-inducible system was cloned into pSB1C3 and co-transformed into DH5-alpha cells along with a plasmid containing heme oxygenase and PcyA. After growing the cells to OD=0.9-1 in Terrific Broth, the cultures were induced with increasing levels of arabinose to study the vibrancy of the cells once harvested (Fig.1).
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12INCOMPATIBLE WITH RFCIllegal NheI site found at 125
- 21INCOMPATIBLE WITH RFCIllegal BamHI site found at 65
Illegal BamHI site found at 731
- 23COMPATIBLE WITH RFC
- 25INCOMPATIBLE WITH RFCIllegal NgoMIV site found at 1251
Illegal AgeI site found at 1341
- 1000COMPATIBLE WITH RFC