Difference between revisions of "Part:BBa K1899009"

Revision as of 21:17, 19 October 2016

B0032-LacI

LacI is conjugated to a medium strength RBS

Construct for Characterisation

Fig.1 lacI-lacp with reporter gene.

In order to characterise the efficiency of this construction intermediate, it was ligated with a construct containing lacp and GFP reporter gene, BBa_E0240. The expression of LacI was driven by a strong constitutive promoter, BBa_J23101, and terminated by BBa_B1006 terminator. The assembly processes are performed in BioBrick RFC10 standard.

Results

Experiments on comparing the GFP expression driven by lacp from constructs with and without BBa_B0032-tetR were performed. Negative control used was BBa_E0240. Results indicated that BBa_B0032-lacI driven by BBa_J23101 and terminated by BBa_B1006 could reduce the GFP expression by xx times.

Fig 2. Comparison on the GFP expression of construct with and without BBa_B0032-tetR. BBa_E0240 was used as negative control. Characterization was done using E. coli strain JW0336. Cells were first precultured overnight and were subcultured to mid-log phase where GFP emission measurements were made using an EnVision® multilabel reader. This result was obtained by combining 3 characterization data obtained in 3 different days. Error bar present SD from 3 biological replicates.

Sequence and Features