Difference between revisions of "Part:BBa K2557007"

 
 
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TP901 is a widely studied recombinase that we use to mediate intracellular signals.
 
TP901 is a widely studied recombinase that we use to mediate intracellular signals.
  
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===Usage and Biology===
 
===Usage and Biology===
  
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TP901 is an recombinase comeing from Lactococcus lactis have been evaluated for their use in mammalian cells. Target DNA sequences can be inserted, excised, inverted, and translocated, and are subject to cassette exchange using these recombinase or integrase systems. The combined or individual use of these systems provides a wide range of control for gene expression.
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In our genetic circuit, TP901 regulated by TetR. When the inhibition released by TEV protease, TP901 can bind to specific attB/P sites to catalyze DNA recombination, leading to the expression of mCherry.
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===Characterization===
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<html>
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<img src="https://static.igem.org/mediawiki/parts/a/a9/T--NAU-CHINA-liushixibao.jpg"width="600"/>
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</html>
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Fig.2 Function verification of reversal efficiency and threshold characteristics of different recombinase in HEK 293 T Cells
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(A)Fluorescence microscope observation of HEK 293T  undergone different experimental treatments
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(B)The statistical chart of average fluorescence intensity of cells shows that the cells with Bxb1 recombinase have a higher fluorescence intensity than those with TP901 recombinase under the same promoter strength and recombinase concentration. However, if the concentration of recombinase is low, there is no significant difference in fluorescence intensity.   
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(C)The statistics of the proportion of fluorescent cells show that the proportion of fluorescent cells has a sudden jump discontinuity between low concentration and high concentration of Bxb1 and TP901 recombinases. Similar results were obtained in all three repetitions.
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The results of image B show that the reverse efficiency of TP901 recombinase is lower than Bxb1 recombinase under the same promoter strength and recombinase concentration. However, if the concentration of recombinase is low, there is no significant difference in fluorescence intensity. The results of the image C show that TP901 recombinases have a threshold property. So, the proportion of fluorescent cells have a jump discontinuity between low concentration and high concentration of recombinase.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2557007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2557007 SequenceAndFeatures</partinfo>

Latest revision as of 18:15, 17 October 2018


TP901 recombinase

TP901 is a widely studied recombinase that we use to mediate intracellular signals.


Usage and Biology

TP901 is an recombinase comeing from Lactococcus lactis have been evaluated for their use in mammalian cells. Target DNA sequences can be inserted, excised, inverted, and translocated, and are subject to cassette exchange using these recombinase or integrase systems. The combined or individual use of these systems provides a wide range of control for gene expression.

In our genetic circuit, TP901 regulated by TetR. When the inhibition released by TEV protease, TP901 can bind to specific attB/P sites to catalyze DNA recombination, leading to the expression of mCherry.

Characterization

Fig.2 Function verification of reversal efficiency and threshold characteristics of different recombinase in HEK 293 T Cells (A)Fluorescence microscope observation of HEK 293T undergone different experimental treatments (B)The statistical chart of average fluorescence intensity of cells shows that the cells with Bxb1 recombinase have a higher fluorescence intensity than those with TP901 recombinase under the same promoter strength and recombinase concentration. However, if the concentration of recombinase is low, there is no significant difference in fluorescence intensity. (C)The statistics of the proportion of fluorescent cells show that the proportion of fluorescent cells has a sudden jump discontinuity between low concentration and high concentration of Bxb1 and TP901 recombinases. Similar results were obtained in all three repetitions.

The results of image B show that the reverse efficiency of TP901 recombinase is lower than Bxb1 recombinase under the same promoter strength and recombinase concentration. However, if the concentration of recombinase is low, there is no significant difference in fluorescence intensity. The results of the image C show that TP901 recombinases have a threshold property. So, the proportion of fluorescent cells have a jump discontinuity between low concentration and high concentration of recombinase.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 633
    Illegal PstI site found at 1344
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 633
    Illegal PstI site found at 1344
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 655
    Illegal BamHI site found at 1599
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 633
    Illegal PstI site found at 1344
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 633
    Illegal PstI site found at 1344
    Illegal NgoMIV site found at 99
    Illegal NgoMIV site found at 243
    Illegal AgeI site found at 6
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 852