Difference between revisions of "Part:BBa K2984025"
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<partinfo>BBa_K2984025 short</partinfo> | <partinfo>BBa_K2984025 short</partinfo> | ||
| − | Another strong constitutive promoter is the AR promoter that is composed of HSP70A and RBCS2 building a tandem promoter. Therefore the HSP70A (A) promoter is fused upstream of another promoter resulting in a stronger transgene expression (Schroda et al. 2000). The combination of the HSP70A (A) promoter and the RBCS2 (R) promoter building < | + | <html> |
| + | This part is a part of the <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>. Another strong constitutive promoter is the AR promoter that is composed of HSP70A and RBCS2 building a tandem promoter. Therefore the HSP70A (A) promoter is fused upstream of another promoter resulting in a stronger transgene expression (Schroda et al. 2000). The combination of the HSP70A (A) promoter and the RBCS2 (R) promoter building <sub>P</sub>AR has the positive effect that transcriptional gene silencing (TGS) is strongly reduced making this promoter popular for transgene expression (Schroda et al. 2002). This part was designed for <i>C. reinhardtii</i>. | ||
| + | </html> | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K2984025 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2984025 SequenceAndFeatures</partinfo> | ||
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| + | == Characterization == | ||
| + | |||
| + | <html> | ||
| + | <figure> | ||
| + | <img src="https://static.igem.org/mediawiki/parts/b/b3/T--Humboldt_Berlin--AR_A1-B1.png" alt="Plate_L0-AR_A1-B1_E.coli" width="500"> | ||
| + | <figcaption>Fig.1 - Successful transformation of L0-AR_A1-B1 into <i>E. coli</i>.</figcaption> | ||
| + | </figure> | ||
| + | </html> | ||
| + | |||
| + | <html> | ||
| + | The AR Promotor was used for two different paromomycin constructs. The first carries the registry part <a href="https://parts.igem.org/Part:BBa_K2703008"> BBa_K2703008 </a> which we wanted to compare to our <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984006"> improved </a> | ||
| + | paromomycin construct which also feautres this promoter. The improvement was done as part of the 2019 <a href="https://2019.igem.org/Judging/Medals"> gold criteria.</a> The colony growth proves the functionality of the promoter. | ||
| + | This part was also used for characterization purposes of the ARS secretion signal in the level 1 construct <a herf="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984033" > L1c-AR-ARS-YFP-Rbcs2 </a> | ||
| + | </html> | ||
| + | <html> | ||
| + | <figure> | ||
| + | <img src="https://2019.igem.org/wiki/images/4/4a/T--Humboldt_Berlin--AR_Paro.jpeg | ||
| + | " width="500"> | ||
| + | <figcaption>Fig.2 - TAP-agar plates containing paromomycin. The <i> C.Reinhardtii </i> colonies growing on the plate were tranformed with our improved <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984006"> paromomycin construct </a> which is flanked by this AR promoter and the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984018"> Rbcs2 </a> terminator </figcaption> | ||
| + | </figure> | ||
| + | </html> | ||
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<partinfo>BBa_K2984025 parameters</partinfo> | <partinfo>BBa_K2984025 parameters</partinfo> | ||
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| + | ==References== | ||
| + | <ol> | ||
| + | <li> | ||
| + | Schroda, M., Blöcker, D., & Beck, C. F. (2000). The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. The plant journal, 21(2), 121-131. | ||
| + | </li> | ||
| + | <li> | ||
| + | Schroda, M., et al. (2002). "Sequence elements within an HSP70 promoter counteract transcriptional transgene silencing in Chlamydomonas." Plant J 31(4): 445-455. | ||
| + | </li> | ||
| + | </ol> | ||
Latest revision as of 21:19, 21 October 2019
AR Promoter A1-B1
This part is a part of the Chlamy-HUB-Collection. Another strong constitutive promoter is the AR promoter that is composed of HSP70A and RBCS2 building a tandem promoter. Therefore the HSP70A (A) promoter is fused upstream of another promoter resulting in a stronger transgene expression (Schroda et al. 2000). The combination of the HSP70A (A) promoter and the RBCS2 (R) promoter building PAR has the positive effect that transcriptional gene silencing (TGS) is strongly reduced making this promoter popular for transgene expression (Schroda et al. 2002). This part was designed for C. reinhardtii.
Usage and Biology
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 282
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterization
The AR Promotor was used for two different paromomycin constructs. The first carries the registry part BBa_K2703008 which we wanted to compare to our improved
paromomycin construct which also feautres this promoter. The improvement was done as part of the 2019 gold criteria. The colony growth proves the functionality of the promoter.
This part was also used for characterization purposes of the ARS secretion signal in the level 1 construct L1c-AR-ARS-YFP-Rbcs2
References
- Schroda, M., Blöcker, D., & Beck, C. F. (2000). The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. The plant journal, 21(2), 121-131.
- Schroda, M., et al. (2002). "Sequence elements within an HSP70 promoter counteract transcriptional transgene silencing in Chlamydomonas." Plant J 31(4): 445-455.