Part:BBa_K4165233
His - UBE2N
This biobrick consists of T7 promotor (BBa_K3633015), RBS (BBa_K4165016), 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.
Source
Synthesized
Usage and Biology
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 328
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Dry lab
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.
WetLab Results
Transformation of His UBE2N in BL-21 using pGS-21a vector
Figure 2. Transformed plate of His UBE2N + pGS-21a
Transformation of His UBE2N in DH-5 alpha using pJET vector
Figure 2. Transformed plate of His UBE2N + pJET
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