Difference between revisions of "Part:BBa K4192110"

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===Characterization===
 
===Characterization===
 
<p>This part has been connected to the downstream of Plac and transferred into <i>E. coli</i> and <i>Pseudomonas fluorescens</i> 2P24 for testing. To get more information about the resut, please see https://parts.igem.org/Part:BBa_K4192114.</p>
 
<p>This part has been connected to the downstream of Plac and transferred into <i>E. coli</i> and <i>Pseudomonas fluorescens</i> 2P24 for testing. To get more information about the resut, please see https://parts.igem.org/Part:BBa_K4192114.</p>
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<p>The results showed that it was difficult to express in <i>E. coli</i>, but it had a good expression effect in <i>Pseudomonas fluorescens</i> 2P24, which means that the two genes can be co-expressed to improve the biofilm production of <i>Pseudomonas fluorescens</i>.</p>
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<p>The data are shown in the figure below:</p>
 +
[[File:CAUChinabiofilmof2P24final.png|600px|thumb|center|]]
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<center><strong> Fig.7 Biofilm of <i>Pseudomonas fluorescens</i> 2P24 </strong></center>
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<p>Through, Levene's test of equality of error vDariances, p<0.05, it is proved that the experimental results of different recombinant bacteria have significant differences.</p>
 +
<p>Through Duncan’s multiple range test, there is little difference between wild type and cdg recombinant bacteria, and there was also little difference between cdg and gacA groups. The biofilm production of cdg+gacA recombinant bacteria iss 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.</p>
 +
<p>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.</p>
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 +
<p>To get more information about the resut, please see https://parts.igem.org/Part:BBa_K4192114.</p>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 07:29, 7 October 2022


cdg-gacA

RBS-cdg-RBS-gacA is composed of cdg gene (BBa_K4192010), gacA gene (BBa_4192011) and strong RBS sequence of Pseudomonas fluorescens. (BBa_K4192000). cdg and gacA encode regulatory factors DGC and GacA in Pseudomonas fluorescens, which can ncrease the expression of extracellular polysaccharide (EPS) and then increase the production of biofilm.

Characterization

This part has been connected to the downstream of Plac and transferred into E. coli and Pseudomonas fluorescens 2P24 for testing. To get more information about the resut, please see https://parts.igem.org/Part:BBa_K4192114.

The data are shown in the figure below:

CAUChinabiofilmof2P24final.png
Fig.7 Biofilm of Pseudomonas fluorescens 2P24

Through, Levene's test of equality of error vDariances, p<0.05, it is proved that the experimental results of different recombinant bacteria have significant differences.

Through Duncan’s multiple range test, there is little difference between wild type and cdg recombinant bacteria, and there was also little difference between cdg and gacA groups. The biofilm production of cdg+gacA recombinant bacteria iss 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.

To get more information about the resut, please see https://parts.igem.org/Part:BBa_K4192114.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 268
    Illegal BamHI site found at 259
    Illegal XhoI site found at 357
    Illegal XhoI site found at 801
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 192
    Illegal NgoMIV site found at 1289
    Illegal AgeI site found at 759
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 210
    Illegal BsaI.rc site found at 777