Team:Tsinghua-E/Result

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Strains

BL21(DE3)(mut-bitter30(+))/pSC101wt-ori-mutD-sfGFP/pTrc99A-trpss-30-tetA

BL21(DE3)(mut-bitter32(+))/pSC101wt-ori-mutD-sfGFP/pTrc99A-trpss-32-tetA

Negative control:

BL21(DE3) pSC101wt-ori-mutD-sfGFP/pTrc99A-trpss-30-tetA

BL21(DE3) pSC101wt-ori-mutD-sfGFP/pTrc99A-trpss-32-tetA

Each evolution was conducted in three parallel with only one parallel of negative control.

Results

By using the parts constructed by us, following the protocol described, we conducted the continuous evolution derived by mutD based mutation for two E. Coli BL21 (DE3) strains carrying bitter defender part 30 and 32. The results were shown below:

Table1 Titer increase dynamics of different strains after different rounds of evolution (rbs30)

 

1st generation

2nd generation

3rd generation

Control (only have mutation part)

No growth

No growth

No growth

Bitter defender part rbs30 (mg/L)

0.791

3.578

3.119

Bitter defender part rbs30/mutation part (mg/L)

0.500
0.741
0.500

 

0.919
3.748

2.245
3.482
2.245

Table2 Titer increase dynamics of different strains after different rounds of evolution (rbs32)

 

1st generation

2nd generation

3rd generation

Control (only have mutation part)

No growth

No growth

No growth

Bitter defender part rbs32 (mg/L)

0.673

1.063

3.433

Bitter defender part rbs32/mutation part (mg/L)

3.142
0.913
0.718

0.766
3.049
4.257

2.540
3.849
3.614