Difference between revisions of "Part:BBa K2149020:Experience"
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Even tough we managed to assemble the Lux Operon, we had not enough time to express it and much less try to biosynthesize our beloved alkanes. So we ended up expressing only the LuxD gene. | Even tough we managed to assemble the Lux Operon, we had not enough time to express it and much less try to biosynthesize our beloved alkanes. So we ended up expressing only the LuxD gene. | ||
We assembled a plasmid with the LuxD gene and a RFP gene to ensure a visual representation of our expression. And the grown bacteria expressed quite a lot of the gene. | We assembled a plasmid with the LuxD gene and a RFP gene to ensure a visual representation of our expression. And the grown bacteria expressed quite a lot of the gene. | ||
| − | + | ||
| + | |||
| + | Pelletized cells | ||
https://static.igem.org/mediawiki/2016/e/e2/T--USP-EEL-Brazil--Bacteria.jpeg | https://static.igem.org/mediawiki/2016/e/e2/T--USP-EEL-Brazil--Bacteria.jpeg | ||
| − | + | Source: Personal archive. | |
So with our visual expression confirmation, we went for the actual confirmation. We then digested our cells and separated our gene from it's plasmid. With our product had both the RFP and the LuxD gene in it's cut product, the relation between the RFP protein and the LuxD protein expressions are then proved. | So with our visual expression confirmation, we went for the actual confirmation. We then digested our cells and separated our gene from it's plasmid. With our product had both the RFP and the LuxD gene in it's cut product, the relation between the RFP protein and the LuxD protein expressions are then proved. | ||
And our gel product proved to be also really intense, with well defined and strong bands. Analyzing the band size, it is also pretty clear the gene composition of it, with around 1740bp for the LuxD gene and 905bp for the RFP, our expression should turn out somewhere between the 2000bp ant 3000bp band, with 2645bp. | And our gel product proved to be also really intense, with well defined and strong bands. Analyzing the band size, it is also pretty clear the gene composition of it, with around 1740bp for the LuxD gene and 905bp for the RFP, our expression should turn out somewhere between the 2000bp ant 3000bp band, with 2645bp. | ||
| + | Electrophoresis from expressed LuxD+RFP, with Quick-Load Purple 2-log DNA ladder | ||
https://static.igem.org/mediawiki/2016/c/cf/T--USP-EEL-Brazil--LuxDGel.jpeg | https://static.igem.org/mediawiki/2016/c/cf/T--USP-EEL-Brazil--LuxDGel.jpeg | ||
| − | + | Source: Personal archive. | |
===User Reviews=== | ===User Reviews=== | ||
Revision as of 16:16, 21 October 2016
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K2149020
Even tough we managed to assemble the Lux Operon, we had not enough time to express it and much less try to biosynthesize our beloved alkanes. So we ended up expressing only the LuxD gene. We assembled a plasmid with the LuxD gene and a RFP gene to ensure a visual representation of our expression. And the grown bacteria expressed quite a lot of the gene.
Pelletized cells
Source: Personal archive.
So with our visual expression confirmation, we went for the actual confirmation. We then digested our cells and separated our gene from it's plasmid. With our product had both the RFP and the LuxD gene in it's cut product, the relation between the RFP protein and the LuxD protein expressions are then proved. And our gel product proved to be also really intense, with well defined and strong bands. Analyzing the band size, it is also pretty clear the gene composition of it, with around 1740bp for the LuxD gene and 905bp for the RFP, our expression should turn out somewhere between the 2000bp ant 3000bp band, with 2645bp.
Electrophoresis from expressed LuxD+RFP, with Quick-Load Purple 2-log DNA ladder
Source: Personal archive.
User Reviews
UNIQ2e7ef32c1ff0fe6a-partinfo-00000000-QINU UNIQ2e7ef32c1ff0fe6a-partinfo-00000001-QINU