Difference between revisions of "Part:BBa K1943001"

 
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<partinfo>BBa_K1943001 short</partinfo>
 
<partinfo>BBa_K1943001 short</partinfo>
  
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Team Uppsala 2012 chromoprotein attracts many interests because it's more convenient than fluorescent protein for it can be observed with naked eyes. However, characterized data of these proteins are few. Because a single coding region is on the plasmid, we constructed BBa_K1943001 with chromoprotein fwYellow(BBa_K1033910) this year and did characterization.
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We successfully constructed eight plasmids with various pigment coding sequence, including eforRed, gfasPurple, Fw Yellow and SpisPink.
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We've constructed a series of plasmids which are all in the same pattern: a strong/weak promoter, a strong/weak RBS and a chromoprotein. We want to monitor the speed of expression of these plasmids in normal incubation conditions (like 37&#8451; overnight for LB agar plate and 37&#8451; 180rpm for LB broth). The expression speed and strength of the constructed biobricks will be carefully monitored and gave others a relative scale for using chromoprotein as a reporter gene.
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Compared to the common used fluorescence protein such as GFP, RFP, chromoprotein is significantly more useful reporter gene. In our parts, the coding sequences of these pigments are all among 700bp long, and the expression efficiency is high according to the experiment results.
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We contributed this series of the chromoprotein plasmid for the reason that they have significant  advantages to be used as backbones. The length of 700bp is easy to distinguish from the original backbone (around 2000bp) by gel electrophoresis when constructing new expression system. Additionally, this kind of backbones are superior because we are able to directly observe the expression results by using naked eyes. It will be much more convenient for the laboratory with limited conditions and also save our time. In the tests, the color of colonies were easy to recognize after 18 hours of incubation.
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Our plasmid can also be a substitute for [[Part:BBa_J04450]], which kindly recommended to be used in plasmid construction by replacing the RFP gene. For the same reason, we provide these pigment sequence to make the construction process more efficiently.
 
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===Usage and Biology===
 
===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1943001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1943001 SequenceAndFeatures</partinfo>
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[[File:FwYellow, yellow chromoprotein reporter system.jpeg|center]]
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Right is BBa_K1943000(Strong Promoter & RBS), Left is BBa_K1943001(Weak Promoter & RBS)
  
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[[File:SUSTech_Pattern_of_chromoprotein_1.jpeg]]
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A few of the chromoproteins form the pattern of S(BBa_K1943000), U(BBa_K1943002), S(BBa_K1943004), T(BBa_K1943006), e(BBa_K1943005), c(BBa_K1943003), h(BBa_K1943001), !(BBa_K1943007)
 
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<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K1943001 parameters</partinfo>
 
<partinfo>BBa_K1943001 parameters</partinfo>
 
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Revision as of 01:19, 10 September 2016


fwYellow, yellow chromoprotein reporter system (medium Promoter , weak RBS)

We successfully constructed eight plasmids with various pigment coding sequence, including eforRed, gfasPurple, Fw Yellow and SpisPink. Compared to the common used fluorescence protein such as GFP, RFP, chromoprotein is significantly more useful reporter gene. In our parts, the coding sequences of these pigments are all among 700bp long, and the expression efficiency is high according to the experiment results. We contributed this series of the chromoprotein plasmid for the reason that they have significant advantages to be used as backbones. The length of 700bp is easy to distinguish from the original backbone (around 2000bp) by gel electrophoresis when constructing new expression system. Additionally, this kind of backbones are superior because we are able to directly observe the expression results by using naked eyes. It will be much more convenient for the laboratory with limited conditions and also save our time. In the tests, the color of colonies were easy to recognize after 18 hours of incubation. Our plasmid can also be a substitute for Part:BBa_J04450, which kindly recommended to be used in plasmid construction by replacing the RFP gene. For the same reason, we provide these pigment sequence to make the construction process more efficiently. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 172
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 549
FwYellow, yellow chromoprotein reporter system.jpeg

Right is BBa_K1943000(Strong Promoter & RBS), Left is BBa_K1943001(Weak Promoter & RBS)

SUSTech Pattern of chromoprotein 1.jpeg A few of the chromoproteins form the pattern of S(BBa_K1943000), U(BBa_K1943002), S(BBa_K1943004), T(BBa_K1943006), e(BBa_K1943005), c(BBa_K1943003), h(BBa_K1943001), !(BBa_K1943007)