Difference between revisions of "Part:BBa K1993014"
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<partinfo>BBa_K1993014 short</partinfo> | <partinfo>BBa_K1993014 short</partinfo> | ||
− | pSMA-eGFP | + | eGFP is a type of GFP derivatives by mutation. This mutation dramatically improved the spectral characteristics of GFP, resulting in increased fluorescence, photostability, and a shift of the major excitation peak to 488 nm, with the peak emission kept at 509 nm. (Details could be seen on [https://parts.igem.org/Part:BBa_K1993017 BBa_K1993017]) |
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+ | Alpha-smooth muscle actin (α-SMA) is the actin isoform that predominates within vascular smooth-muscle cells and plays an important role in fibrogenesis. (Details could be seen on [https://parts.igem.org/Part:BBa_K1993022 BBa_K1993022]) | ||
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+ | Previous studies indicated that alpha-smooth muscle actin (α-SMA) could be induced in inflammatory environment and had a relationship with the risk of MSC fibrogenesis, which weakened their therapeutic effect. [1] As a result, we desired to find out a special way to detect the expression of α-SMA in MSCs. | ||
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+ | To achieve it, we designed a plasmid with promoter of SMA (pSMA) linking eGFP. (Figure 1) | ||
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+ | <html><p align="center"> | ||
+ | <img src="https://static.igem.org/mediawiki/2016/2/29/T--SYSU-MEDICINE--pSMA-eGFP.png" style="width:400px" ></a> | ||
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+ | </p></html> <br> | ||
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+ | <p align="center">'''Figure 1 pSMA-eGFP'''</p><br> | ||
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+ | To confirm the function of this plasmid, we administered TGF-β1 (7ng/mL). After that, Promoter of α-SMA was activated and green fluorescence could be detected in MSCs (Figure 2) | ||
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+ | <html><p align="center"> | ||
+ | <img src="https://static.igem.org/mediawiki/2016/4/42/T--SYSU-MEDICINE--BBa_K1993014-fig2.png" style="width:300px" ></a> | ||
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+ | </p></html> <br> | ||
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+ | <p align="center">'''Figure 2 Promoter of α-SMA was activated and green fluorescence could be detected'''</p><br> | ||
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+ | ==Reference== | ||
+ | [1] Talele N P, Fradette J, Davies J E, et al. Expression of α-smooth muscle actin determines the fate of mesenchymal stromal cells[J]. Stem cell reports, 2015, 4(6): 1016-1030. | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Latest revision as of 12:48, 18 October 2016
pSMA-eGFP
eGFP is a type of GFP derivatives by mutation. This mutation dramatically improved the spectral characteristics of GFP, resulting in increased fluorescence, photostability, and a shift of the major excitation peak to 488 nm, with the peak emission kept at 509 nm. (Details could be seen on BBa_K1993017)
Alpha-smooth muscle actin (α-SMA) is the actin isoform that predominates within vascular smooth-muscle cells and plays an important role in fibrogenesis. (Details could be seen on BBa_K1993022)
Previous studies indicated that alpha-smooth muscle actin (α-SMA) could be induced in inflammatory environment and had a relationship with the risk of MSC fibrogenesis, which weakened their therapeutic effect. [1] As a result, we desired to find out a special way to detect the expression of α-SMA in MSCs.
To achieve it, we designed a plasmid with promoter of SMA (pSMA) linking eGFP. (Figure 1)
Figure 1 pSMA-eGFP
To confirm the function of this plasmid, we administered TGF-β1 (7ng/mL). After that, Promoter of α-SMA was activated and green fluorescence could be detected in MSCs (Figure 2)
Figure 2 Promoter of α-SMA was activated and green fluorescence could be detected
Reference
[1] Talele N P, Fradette J, Davies J E, et al. Expression of α-smooth muscle actin determines the fate of mesenchymal stromal cells[J]. Stem cell reports, 2015, 4(6): 1016-1030.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1763
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 887