Difference between revisions of "Part:BBa K314104"

Revision as of 20:02, 24 October 2010

T7 expression cassette

Lactose (IPTG) inducible T7 protein expression insert includes f1 origin BBa_K314110, a Lac I generator BBa_K314111, a T7 promoter BBa_K314112, and the Elowitz standard RBS BBa_B0034. When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.

Usage and Biology

BBa_E0040 (GFP) was inserted between S and P using standard BioBrick assembly. The resulting constructs were grow in DH5a (constitutive promoters BBa_J23100 or BBa_J23114, as well as inducible promoter BBa_R0011) or BL21(DE3)* (T7 promoter, BBa_K314104) cells overnight at 298K in either the presence or absence of 0.5mM IPTG. The cells were harvested, washed in PBS, and then fluorescence for the whole cells was measured on a fluorometer (ex. 485, em. 525) and normalized to cell density (OD600). Cells that did not contain GFP were used as a blank. As observed, the addition of IPTG significantly increased the observed fluorescence of the cells.

Sequence and Features