Difference between revisions of "Part:BBa K2549044"
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| + | ==Experimental Characterization by AFCM-Egypt== | ||
| + | In order to amplify this DNA part, we used PCR amplification to reach the desired concentration to complete our experiments using specific forward and reverse primers, running the parts on gel electrophoresis as this part presents in lane (P1) including CD8 alpha-his tag-mouse notch core-ZF21.16\VP64, and then measuring the specific concentration of the running part using Real-Time PCR as shown in the following figure. | ||
| + | <html><div align="center"style="border:solid #17252A; width:80%;float:center;"><img style=" max-width:850px; | ||
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| + | "src="https://static.igem.wiki/teams/4586/wiki/parts-experiments/pcr-ampli.png"> | ||
| + | <p class=MsoNormal align=center style='text-align:left;border:none;width:98% ;justify-content:center;'><span | ||
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| + | We performed the double digestion method for this part in the prefix and suffix with its specific restriction enzyme and applied this part to gel electrophoresis as shown in the following figure in lane (P1) | ||
| + | <html><div align="center"style="border:solid #17252A; width:80%;float:center;"><img style=" max-width:850px; | ||
| + | width:100%; | ||
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| + | "src="https://static.igem.wiki/teams/4586/wiki/parts-experiments/digestion-2.png"> | ||
| + | <p class=MsoNormal align=center style='text-align:left;border:none;width:98% ;justify-content:center;'><span | ||
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<!-- 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 20:50, 11 October 2023
CD8alpha signal peptide
CD8α signal peptide guides synthesized fusion protein to pass the translocon[1] into the endoplasmic reticulum[2], and the fusion protein will be later sugar modified in Golgi[3], presented on the plasma membrane and located to the outside of the cell. We include this part as it is required for the parts collection Part:BBa_K2549016 ~ Part:BBa_K2549043.
Literature Characterization by AFCM-Egypt
The study created a reporter construct by joining the C-terminus of CD63, one of the most used exosome markers, to nanoluc (nluc), a tiny and potent bioluminescence reporter10. After progressive centrifugation to eliminate masking signals12, luminescence in the cell-culture supernatant was measured. This reporter gene was co-transfected with plasmids expressing potential candidates for exosome production augmentation.
Poly(A) RNA samples from each cell line were separated by gel electrophoresis, transferred to a nitrocellulose membrane, and hybridized with 32P-labeled cDNA probes specific for MC-CPA or CD8amp;. The membranes were then reprobed with a 32P-labeled p-actin cDNA probe to verify that equal amounts of RNA had been loaded in each lane. The results showed that MC-CPA mRNA was expressed at a higher level in MW9.11-3 cells than in MC/9.IL-4 cells, while CDalpha mRNA was expressed at a similar level in both cell lines. These findings suggest that MC-CPA and CDalpha may play different roles in the development and differentiation of these two cell
Experimental Characterization by AFCM-Egypt
In order to amplify this DNA part, we used PCR amplification to reach the desired concentration to complete our experiments using specific forward and reverse primers, running the parts on gel electrophoresis as this part presents in lane (P1) including CD8 alpha-his tag-mouse notch core-ZF21.16\VP64, and then measuring the specific concentration of the running part using Real-Time PCR as shown in the following figure.
We performed the double digestion method for this part in the prefix and suffix with its specific restriction enzyme and applied this part to gel electrophoresis as shown in the following figure in lane (P1)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]