Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Casey Martin
Date created: 2014-10-16 11:00:00
Date modified: 2015-05-08 01:10:23
Plasmid producing corn NAAT1, DMAS, and TOM1
| Types | DnaRegion |
| Roles | plasmid Plasmid |
| Sequences | BBa_K1422506_sequence (Version 1) |
Description
This plasmid codes for NAAT1 (BBa_K1422502) and DMAS (BBa_K1422503), two of the four enzymes used to produce corn phytosiderophores; TOM1 (BBa_K1422507), which transports phytosiderophores out of cells; and sfGFP, as a reporter. BBa_K1422505 goes along with this plasmid, producing the other two enzymes necessary for the production of corn phytosiderophores, and includes a killswitch.Notes
Using a conditional promoter so that the modified Bacillus subtilis would only produce phytosiderophores in the presence of plants roots was considered, but it was decided that conditional promoters would not be reliable enough. A constitutive promoter of medium strength was chosen for the corn genes because a weak promoter might not have produced enough phytosiderophores to make a difference, and a strong promoter might have overtaxed the cell in the production of phytosiderophores as to be counterproductive. The reporter is placed after the corn genes because gene expression decreases linearly as the distance from the promoter increases, and expressing the corn genes is much more important than expressing the reporter. sfGFP was used as a reporter because it is easily observed, easily measured, and expressed in Bacillus subtilis. The RBS SpoVG was chosen for its relative strength in Bacillus subtilis. A relatively strong terminator with few base pairs was selected for its relative effectiveness and low number of base pairs. NAAT can be coded by naat-A, naat-B, or naat-1; naat1 from corn was selected because a previous experiment revealed that the rice naat1 gene encodes functional NAAT and is expressed in cells of Fe-deficient leaves, strongly suggesting that it is involved in phytosiderophores production and sufficient to produce NAAT on its own, without other naat genes.Source
NAAT1:NCBI. (19 October, 2011). Zea mays ZmNAAT1 mRNA for putative nicotianamine aminotransferase, complete cds. http://www.ncbi.nlm.nih.gov/nuccore/166788521
DMAS:
NCBI. (2014). deoxymugineic acid synthase1 [Zea mays]. http://www.ncbi.nlm.nih.gov/protein/162460852
TOM1:
NCBI. (10 December, 2008). Zea mays clone 222622 protein transporter mRNA, complete cds. http://www.ncbi.nlm.nih.gov/nuccore/195621011
| Access | Instance | Definition |
| public public public public public public public public public public | BBa_K143012 BBa_K143021 BBa_K1422502 BBa_K143021 BBa_K1422503 BBa_K143021 BBa_K1422507 BBa_K143021 BBa_I746916 BBa_B1006 | Pveg RBS-spoVG BBa_K1422502 RBS-spoVG BBa_K1422503 RBS-spoVG BBa_K1422507 RBS-spoVG BBa_I746916 BBa_B1006 |