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Team:Stanford-Brown/HellCell/Plasmid
From 2012.igem.org
(→The Test Plasmid) |
(→The Test Plasmid) |
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[[File:TestPlasmid1.png|400px|center]] | [[File:TestPlasmid1.png|400px|center]] | ||
| - | Figure 1: Test Plasmid, inserting 1 gene | + | '''Figure 1: Test Plasmid, inserting 1 gene |
P = Strong Promoter, BBa_J23100 | P = Strong Promoter, BBa_J23100 | ||
R = Strong RBS, BBa_B0030 | R = Strong RBS, BBa_B0030 | ||
G = Gene of interest | G = Gene of interest | ||
| - | T = GFP Reporter, BBa_E0840 | + | T = GFP Reporter, BBa_E0840''' |
In some cases, we wished to insert 2 genes into the same plasmid as they both contributed to one pathway. We used the following scheme: | In some cases, we wished to insert 2 genes into the same plasmid as they both contributed to one pathway. We used the following scheme: | ||
[[File:TestPlasmid2.png|400px|center]] | [[File:TestPlasmid2.png|400px|center]] | ||
| - | Figure 2: Test Plasmid, inserting two-gene pathway | + | '''Figure 2: Test Plasmid, inserting two-gene pathway |
P = Strong Promoter, BBa_J23100 | P = Strong Promoter, BBa_J23100 | ||
R = Strong RBS, BBa_B0030 | R = Strong RBS, BBa_B0030 | ||
| - | G1 = Gene of interest, | + | G1 = Gene of interest, first step of pathway |
| - | G2 = Gene of interest, | + | G2 = Gene of interest, second step of pathway |
| - | T = GFP Reporter, BBa_E0840 | + | T = GFP Reporter, BBa_E0840''' |
Revision as of 10:27, 3 October 2012
The Test Plasmid
To test the genes that we isolated from various organisms in hopes of arming our Escherichia coli from the extremes, we used standard BioBrick assembly to place all of the genes into the same "Test Plasmid." We used a strong promoter and RBS in hopes of producing as much of the proteins as possible in the cell, increasing the chances of observing their function. Instead of a regular terminator, we used GFP-producing part BBa_E0840 so that we could have a rough, easy visual confirmation that our protein is probably being transcribed (See Figure 1). Because of the way that we assembled the parts together, all of these composite parts ended up on psb1A2 (the plasmid BBa_E0840 was on).
Figure 1: Test Plasmid, inserting 1 gene P = Strong Promoter, BBa_J23100 R = Strong RBS, BBa_B0030 G = Gene of interest T = GFP Reporter, BBa_E0840
In some cases, we wished to insert 2 genes into the same plasmid as they both contributed to one pathway. We used the following scheme:
Figure 2: Test Plasmid, inserting two-gene pathway P = Strong Promoter, BBa_J23100 R = Strong RBS, BBa_B0030 G1 = Gene of interest, first step of pathway G2 = Gene of interest, second step of pathway T = GFP Reporter, BBa_E0840
