Difference between revisions of "Part:BBa K4165233"
 
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This biobrick consists of T7 promotor (BBa_K3633015), RBS (BBa_K4165075), His 6-His tag (BBa_K4165020), UBE2N (BBa_K4165011), T7 Terminator (BBa_K731721), The His tag was attached to the UBE2N coding sequence to serve in the purification using NI-NTA column.
 
This biobrick consists of T7 promotor (BBa_K3633015), RBS (BBa_K4165075), His 6-His tag (BBa_K4165020), UBE2N (BBa_K4165011), T7 Terminator (BBa_K731721), The His tag was attached to the UBE2N coding sequence to serve in the purification using NI-NTA column.
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===Source===
 
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Synthesized
 
  
 
===Usage and Biology===
 
===Usage and Biology===
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this part is a Ubiquitin-conjugating E2 ligase which has a role in the ubiquitination cascade for protein degradation. It has been used in this project to target and degrade both tau and Aβ proteins which are both considered the main causes of Alzheimer’s Disease pathogenesis.
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This part encodes for the UBE2N enzyme which is a member of the E2 ubiquitin-conjugating enzyme family. It acts as the second enzyme in the ubiquitination cascade to receive the thio-esterified ubiquitin from the E1 active site. it will bind to the His tag For the purification of the protein.
  
 
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<partinfo>BBa_K4165233 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4165233 SequenceAndFeatures</partinfo>
  
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===Dry lab===
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===Mathematical Modeling===
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<p style=" font-weight: bold; font-size:14px;"> Mathematical modeling </p>
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<p style=" font-weight: bold; font-size:14px;">Transcription rate and translation rate under T7 promotor </p>
 
<p style=" font-weight: bold; font-size:14px;">Transcription rate and translation rate under T7 promotor </p>
 
the mathematical modeling was based on our code for the calculation of transcription and translation (you can find it in the code section) beside with the estimated results from the wet lab.  
 
the mathematical modeling was based on our code for the calculation of transcription and translation (you can find it in the code section) beside with the estimated results from the wet lab.  
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                     Figure 1. this figure shows the results from the transcription and translation code showing the  
 
                     Figure 1. this figure shows the results from the transcription and translation code showing the  
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                            variation of mRNA and protein concentrations with time.
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                                      variation of mRNA and protein concentrations with time.
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===WetLab Results===
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<p style=" font-weight: bold; font-size:14px;"> Transformation of His UBE2N in BL-21 using pGS-21a vector </p>
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<html>
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<p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/wetlab-results/ube2n-pgs.jpg" style="margin-left:200px;" alt="" width="500" /></p>
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</html>
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                                  Figure 2. Transformed plate of His UBE2N + pGS-21a
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<p style=" font-weight: bold; font-size:14px;"> Transformation of His UBE2N in DH-5 alpha using pJET vector </p>
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<html>
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<p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/wetlab-results/ube2n-pjet.jpg" style="margin-left:200px;" alt="" width="500" /></p>
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</html>
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                                Figure 2. Transformed plate of His UBE2N + pJET
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<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K4165233 parameters</partinfo>
 
<partinfo>BBa_K4165233 parameters</partinfo>
 
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Latest revision as of 12:17, 12 October 2022


His - UBE2N

This biobrick consists of T7 promotor (BBa_K3633015), RBS (BBa_K4165075), His 6-His tag (BBa_K4165020), UBE2N (BBa_K4165011), T7 Terminator (BBa_K731721), The His tag was attached to the UBE2N coding sequence to serve in the purification using NI-NTA column.

Usage and Biology

This part encodes for the UBE2N enzyme which is a member of the E2 ubiquitin-conjugating enzyme family. It acts as the second enzyme in the ubiquitination cascade to receive the thio-esterified ubiquitin from the E1 active site. it will bind to the His tag For the purification of the protein.

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 328
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Mathematical Modeling

Transcription rate and translation rate under T7 promotor

the mathematical modeling was based on our code for the calculation of transcription and translation (you can find it in the code section) beside with the estimated results from the wet lab. 

                    Figure 1. this figure shows the results from the transcription and translation code showing the 
                                     variation of mRNA and protein concentrations with time.