Difference between revisions of "Part:BBa K2371012"

 
 
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<partinfo>BBa_K2371012 short</partinfo>
 
<partinfo>BBa_K2371012 short</partinfo>
  
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This chromoprotein eforRed naturally exhibits red/pink color when expressed. The color is slightly weaker than RFP. On agar plates and in liquid culture, the color is readily visible to naked eye in less than 24 hours of incubation. The DNA was codon-optimized for expression in E.coli and synthesized by the Korean company Bioneer Corporation.
 
  
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In cell and molecular biology, the GFP gene is often used as reporter.
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We add a T7 promotor and a RBS in front of the GFP in order to improve it to be utilized in our paired dCas9 T7 report system in cell free system together with N-t7-dCas9(BBa_K2371000) and C-t7-dCas9(BBa_K2371001).
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The user need to conduct a PCR to linearize the part before reacting with T7 report system.
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We conducted both PCR check(VF and VR) and enzyme check(XbaI and SpeI)
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[[File:BGIC-Union Part 123 GFP.png|700px]]
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<h5>Figure 1. The plasmid of pT7-amilGFP. pT7-amilGFP is composed of a T7 promotor, an RBS and the coding sequence of GFP. This plasmid is used to act as report gene in our report system.</h5>
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[[File:Pcr test of eforRed and GFP.png|400px]]
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<h5>Figure 2. PCR check of eforRed and amliGFP. The expected result of eforRed should be around 1000bp. The insufficient specificity of primers produce another band but the band around 1000bp can present the success of construction. The expected result of amliGFP should be around 1000bp. The control is J23119 biobrick.</h5>
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[[File:RD test of amliRed and GFP.png|400px]]
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<h5>Figure 3. RD check of eforRed and amliGFP. XbaI and SpeI were used. The expected length of both amliGFP and eforRed should be around 750bp.</h5>
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 20:00, 1 November 2017


pT7-amilGFP


In cell and molecular biology, the GFP gene is often used as reporter. We add a T7 promotor and a RBS in front of the GFP in order to improve it to be utilized in our paired dCas9 T7 report system in cell free system together with N-t7-dCas9(BBa_K2371000) and C-t7-dCas9(BBa_K2371001).

The user need to conduct a PCR to linearize the part before reacting with T7 report system.

We conducted both PCR check(VF and VR) and enzyme check(XbaI and SpeI)

BGIC-Union Part 123 GFP.png

Figure 1. The plasmid of pT7-amilGFP. pT7-amilGFP is composed of a T7 promotor, an RBS and the coding sequence of GFP. This plasmid is used to act as report gene in our report system.

Pcr test of eforRed and GFP.png

Figure 2. PCR check of eforRed and amliGFP. The expected result of eforRed should be around 1000bp. The insufficient specificity of primers produce another band but the band around 1000bp can present the success of construction. The expected result of amliGFP should be around 1000bp. The control is J23119 biobrick.


RD test of amliRed and GFP.png

Figure 3. RD check of eforRed and amliGFP. XbaI and SpeI were used. The expected length of both amliGFP and eforRed should be around 750bp.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 600
    Illegal AgeI site found at 712
  • 1000
    COMPATIBLE WITH RFC[1000]