Team:BFSUICC-China/Experiments


BFSUICC-China

 

 

 

 

 

 

 

 

Test the function of parts and circuit

PROTEIN EXPRESSION MEASUREMENTS

In order to show that our synthetic bacteria have a fully functional part that works, expression measurements were made. The construct was usually linked to superfold GFP. The fluorescent intensity can be measured in different ways and is directly related to the expression frequency of the construct (the more intense the signal the more expression you have). For most parts plate reader and microscopy experiments were done.

Plate Reading (Relative and Absolute Fluorescence)

A plate reader is able to take optical density (OD600nm) and fluorescent measurements over time. OD is a measure of bacterial growth over time and fluorescence a measure of protein expression over time.

(I)Comparison between BBa_K1555000 and BBa_K2555000

Plasmid: PSB4K5

Chassis: E.coli Top 10

At 0，1，2，3，4，5，6，7，8，9 and 10h, fluorescent intensity and OD600 were measured 4-48 well plates

(II) BBa_2555003 pBAD-RBS-sfGFP

At 0，1，2，3，4，5，6，7，8，9 and 10h measure fluorescent intensity and OD600 by plate reading

2 48-well plates

(III)BBa_2555004 pCopA-RiboJ-RBS-sfGFP+pBAD-RBS-CueR

Bacteria with o, o.1, o.4, 1.6, 6.4 and 12.8 mM arabinose. At 0，5 and 10h fluorescent intensity and OD600 were measured.

2 48-well plates

（IV）For modeling and real world

BBa_2555004 pCopA-RiboJ-RBS-sfGFP+pBAD-RBS-CueR(0.1mM Arabinose)

Bacteria with o.1 mM arabinose. At 0，5 h fluorescent intensity and OD600 were measured.