"
Team:DTU-Denmark/HelloWorld
From 2013.igem.org
(→Results) |
|||
| Line 8: | Line 8: | ||
Since we are working with many periplasmic proteins, we wanted to try to target proteins to the periplasm. To do this, we used periplasmic signal peptides from the TAT and Sec pathways, and with a translational fusion of the signal peptide to GFP, we expressed GFP in the periplasm. Simultaneously, we expressed RFP in the cytoplasm. | Since we are working with many periplasmic proteins, we wanted to try to target proteins to the periplasm. To do this, we used periplasmic signal peptides from the TAT and Sec pathways, and with a translational fusion of the signal peptide to GFP, we expressed GFP in the periplasm. Simultaneously, we expressed RFP in the cytoplasm. | ||
| - | |||
| - | |||
| + | == Methods == | ||
| + | |||
| + | We followed this protocol to [https://2013.igem.org/Team:DTU-Denmark/Methods/Visualizing_GFP_in_the_periplasm visualize GFP in the periplasm]. | ||
| + | |||
| + | == Results == | ||
[[File:GFP in perimplasm RFP in cytoplasm.png|left|thumb|400px|Transformed ''E. coli'' taken with a confocal microscope]] | [[File:GFP in perimplasm RFP in cytoplasm.png|left|thumb|400px|Transformed ''E. coli'' taken with a confocal microscope]] | ||
[[File:GFP in perimplasm RFP in cytoplasm close up.png|left|thumb|400px|Close up of several cells, showing GFP expression in the periplasm]] | [[File:GFP in perimplasm RFP in cytoplasm close up.png|left|thumb|400px|Close up of several cells, showing GFP expression in the periplasm]] | ||
[[File:Graf.PNG|left|thumbnail|400px|Fluorescence spectrum cross section of one transformed ''E. coli'' cell]] | [[File:Graf.PNG|left|thumbnail|400px|Fluorescence spectrum cross section of one transformed ''E. coli'' cell]] | ||
| - | |||
| - | |||
== Conclusions == | == Conclusions == | ||
Revision as of 13:28, 29 September 2013
Hello World Pilot Project
Contents |
Introduction
‘Hello World!’ are the first words a programmer prints when learning a new programming language. In analogy to this our team decided to do a ‘Hello World’ project in order to familiarize ourselves with lab techniques that we used later on to construct plasmids. Specifically we were performing PCR with uracil-containing primers, purifying PCR products and ligating them by means of USER cloning.
Since we are working with many periplasmic proteins, we wanted to try to target proteins to the periplasm. To do this, we used periplasmic signal peptides from the TAT and Sec pathways, and with a translational fusion of the signal peptide to GFP, we expressed GFP in the periplasm. Simultaneously, we expressed RFP in the cytoplasm.
Methods
We followed this protocol to visualize GFP in the periplasm.
Results
Conclusions
Biobrick BBa_K1067009 successfully directs proteins to the periplasm in E. coli.
