Difference between revisions of "Part:BBa K3121003"
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https://static.igem.org/mediawiki/parts/1/19/T--IISERB_Bhopal--Cpn_10%2B60_lowppx.png | https://static.igem.org/mediawiki/parts/1/19/T--IISERB_Bhopal--Cpn_10%2B60_lowppx.png | ||
| − | <b> Characterization </b> | + | <b> Characterization </b><br> |
| − | We obtained the Cpn 10 /60 by sequential cloning of Cpn 60 and Cpn 10 into pUC18 plasmid (in that particular order).<br> | + | <p>We obtained the Cpn 10 /60 by sequential cloning of Cpn 60 and Cpn 10 into pUC18 plasmid (in that particular order).<br> |
https://2019.igem.org/wiki/images/3/3c/T--IISER_Bhopal--cpn1060result.png<br> | https://2019.igem.org/wiki/images/3/3c/T--IISER_Bhopal--cpn1060result.png<br> | ||
| − | Similar to GroEL/GroES, they function by associating with each other so as to enable better folding of proteins. We characterized the composite part and checked how its induction affected cell growth. The results of the growth analysis may be seen in the graphs provided below. The temperatures chosen were 37°C (as a model for optimal conditions) and 18°C (as a model for suboptimal conditions). While we ideally would have liked to achieve even lower temperatures (as our model clearly predicts that TF could survive near 0°C-like conditions), we were most unfortunately limited by the available infrastructural facilities. | + | Similar to GroEL/GroES, they function by associating with each other so as to enable better folding of proteins. We characterized the composite part and checked how its induction affected cell growth. The results of the growth analysis may be seen in the graphs provided below. The temperatures chosen were 37°C (as a model for optimal conditions) and 18°C (as a model for suboptimal conditions). While we ideally would have liked to achieve even lower temperatures (as our model clearly predicts that TF could survive near 0°C-like conditions), we were most unfortunately limited by the available infrastructural facilities.</p> |
https://static.igem.org/mediawiki/parts/2/29/60_g1_lowppx.png | https://static.igem.org/mediawiki/parts/2/29/60_g1_lowppx.png | ||
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https://static.igem.org/mediawiki/parts/7/7a/60_g2_lowppx.png | https://static.igem.org/mediawiki/parts/7/7a/60_g2_lowppx.png | ||
| − | <b>Inference</b> | + | <b>Inference</b><br> |
| − | From the above graphs, we see that cells induced with Cpn10/60 were able to thrive in both optimal as well as | + | From the above graphs, we see that cells induced with Cpn10/60 were able to thrive in both optimal as well as sub-optimal condition. This could be because of the over expression of the second set of chaperones allows for greater protein folding. |
Latest revision as of 20:21, 21 October 2019
RBS_Cpn10+RBS_Cpn60
We identified the Cpn 10/60 chaperone system (isolated from Oleispira antarctica). The chaperonin 10/60 is homologous to the GroEL/ES chaperonin system in E. coli. Essentially, these are active assistants for protein folding processes. The advantage here is that Cpn 10/60 is a cold-active component, i.e., unlike the mesophilic chaperone systems which are highly inefficient at lower temperatures, these cold-active entities perform optimally at cold temperatures. The Cpn 10/60 which predominantly retains a double ring form at mesophilic temperature ranges (excess of 24℃), shifts to a single ring form at lower temperatures (4-10℃). This is especially relevant since single ring conformations are more energetically efficient.
Characterization
We obtained the Cpn 10 /60 by sequential cloning of Cpn 60 and Cpn 10 into pUC18 plasmid (in that particular order).
Similar to GroEL/GroES, they function by associating with each other so as to enable better folding of proteins. We characterized the composite part and checked how its induction affected cell growth. The results of the growth analysis may be seen in the graphs provided below. The temperatures chosen were 37°C (as a model for optimal conditions) and 18°C (as a model for suboptimal conditions). While we ideally would have liked to achieve even lower temperatures (as our model clearly predicts that TF could survive near 0°C-like conditions), we were most unfortunately limited by the available infrastructural facilities.
Inference
From the above graphs, we see that cells induced with Cpn10/60 were able to thrive in both optimal as well as sub-optimal condition. This could be because of the over expression of the second set of chaperones allows for greater protein folding.