Difference between revisions of "Part:BBa K1916100"

Revision as of 06:56, 25 October 2016

Ara-Inducible NpF2164g5 Expression System

A GAF domain of the cyanobacteriochrome (CBCR) NpF2164 with an intein/chitin-binding domain fused to the C-terminus for purification. Expression regulated by a pBAD promoter. As with most CBCRs, NpF requires a phycocyanobilin co-factor, and must therefore be expressed in a system with heme oxygenase (BBa_I15008) and PcyA (BBa_I15009). NpF2164g5 expresses well in E. coli and cells appear vibrant blue-green under normal light.

Characterization

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 Pellets of cells expressing NpF, spun down 4 hrs after being induced by different concentrations of arabinose.

Sequence and Features