Part:BBa_K4192112
Plac-RBS-gacA
This part is used to test the role of gacA (BBa_K4192011) in biofilm production. We used IPTG to induce its expression, and then used crystal violet to dye 96 well plates. The results are shown in the figure below.
It can be seen that the production of biofilm increased significantly, but the effect of this part is not stable, and further repeated experiments are needed.
Characterization
We try to increase the expression number of gacA gene in E.coli and Pseudomonas fluorescens 2P24 and test its role in increasing biofilm production
We constructed the part BBa_K4192112 and used crystal violet staining to quantitatively test its role in biofilm production. To get more information, please see https://parts.igem.org/Part:BBa_K4192114.
In E.coli, there is no significant difference between engineered bacteria and wild type, but, it shows obvious difference in Pseudomonas fluorescens 2P24.
Through Duncan’s multiple range test, there is little difference between cdg and gacA groups. The biofilm production of cdg and gacA recombinant bacteria is significantly higher than that of the other two groups. This shows that the co-expression effect of cdg and gacA genes is better than that of the two genes alone. The analysis shows that both cdg and gacA genes can increase the concentration of c-di-GMP in bacteria, co-expression can make the concentration of c-di-GMP exceed the threshold and regulate the production of a large number of biofilms.
We can draw a conclusion that increasing the copy number of cdg and gacA at the same time can significantly increase the production of biofilm. Related parallel experiments further verify our conclusion. However, the effect of separate expression of two genes still needs to be determined by further experiments.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 521
- 1000COMPATIBLE WITH RFC[1000]
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