Part:BBa_K2606005:Design
anti_acid and anti_cold part
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 954
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
It is the final composite biobrick of our project, we overexpress anti-acid gene msmK and anti-cold gene cspD2 via the NICE system.
Usage and biology
The gene cspD2 was ligated to the Pnz8149/msmK plasmid by one-step cloning (seamless ligation), and the recombinant plasmid was introduced into the constructed L. lactis NZ3900/pNZ8149-msmk-cspD2 strain using electroporation.
The gfp gene was inserted as a marker gene, and cell viability was characterized by fluorescence intensity. The strain was tested for acid resistance and freezing resistance using a flow cytometer. The process of acid stress and cold stress is similar with the above demonstration process.
Acid stress
Deal with the samples under pH 4.0 with nisin as an inducer.
Fig 1 Number of colonies at acid stress (pH 4.0).
Fig 2 Survival rate at acid stress (pH4.0).
Fig 3 The Comparison curve of survival rate under cold stress.
After 4 consecutive repeated freeze-thaw tests, the recombinant strain was 47.5 times more viable than the control strain, indicating the antifreeze survival rate of the strain with increased overexpression of msmK-cspD2.
Source
The source of the part will be its components.