Team:XMU-China/Parts1

Table 1. The Bio-bricks used in the study

This plasmid consists of two parts of target genes: gfp and luxI, and both of them are regulated by quorum sensing promoter. High copy number. (Fig.7-1)

Fig.7-1 AGE confirmation of plasmid pSB1C3-gfp-luxI construction

This plasmid consists of two parts of target genes: sfgfp and luxI, and both of them are regulated by quorum sensing promoter. sfGFP gene is included into this plasmid as a parallel experiment group of pSB1C3-gfp-luxI. High copy number. (Fig.7-2)

Fig.7-2 AGE confirmation of plasmid pSB1C3-sfgfp-luxI construction

This plasmid consists of two parts of target genes: gfp and luxI, and both of them are regulated by quorum sensing promoter. Middle copy number. (Fig.7-3)

Fig.7-3 AGE of p3H-gfp construction

This plasmid consists of two parts of target genes: sfgfp and luxI, and both of them are regulated by quorum sensing promoter. sfGFP gene is included into this plasmid as a parallel experiment group of pSB1C3-gfp-luxI. Middle copy number. (Fig.7-4)

Fig.7-4 AGE of p3H-sfgfp construction

This plasmid has one target gene aiiA under the regulation of quorum sensing promoter. Middle copy number. (Fig. 7-5)

Fig.7-5 AGE confirmation of plasmid pSB3T5-aiiA construction

This plasmid has one target gene aiiA under the regulation of quorum sensing promoter. Low copy number. (Fig. 7-6)

Fig.7-6 AGE confirmation of plasmid pSB4K5-ndh construction

SDS-PAGE Confirmation for Plasmids

Please refer to Improvement (For a Better Glee) in Strains comparisons.

Future Work (Keep Practicing)

Oh, no, time is up for iGEM Competition, our glee has no more time to play, but pack their luggage and head for MIT. But let's see what they have made so far and what they still have to do if they really want to be a success.

Construction Have done:

Need to do:

Microfluidic

Have done:Finished building two different arrays, find basic testing parameters.

To do:Confirm more suitable microfluidic parameters for oscillation.

Strain

Have done:Confirmed DH5a's incompetent in express oscillation;

To do:Make a comparison between BL21 (WT) and BL21 (DE3) in SDS-PAGE and so on.

Copy number

Have done:Have done: Compared the arrangements of high-middle-low copy number and middle-middle-low copy number's effect on oscillation.

To do:Try other arrangements..

Fast Degrading Tag

Have done:Found LVA-tag is not fast enough for our oscillation circuit.

To do:Try LAA or other fast degrading tag for comparison.