Session with our advisor Miguel A.
• changes in pH due to colorimetry or electrodes, choose the electrodes wisely.
• Nickel transporter in E.coli. * if they send the mutated strain: Transcriptional merger with the trp operon; induce the repressor off.
• Introducing the gene into a multicopy plasmid and select controls (-) and (+)
• Coli introduces at least 40% less zinc when it has the gene. While the flow is still measurable there is no problem.
• An electrode is not really necessary, this can be done with conventional methods.
Research: Efflux pumps of metals.
(It must be ionized, maybe a simple salt, the anion doesn't matter).
Team work:
• Cobalt - Mariana
• Zinc - Jimena
• Cadmium - Chicken
• Nickel - Libertad
• Iron - Atahualpa
• TeO3 (+2) - Isaac
• Cu (2 +) - Minerva
• CrO4 (2 -) - Daniela
• AsO4 (3 -) - Enrique
• Hg - Carlos
• Pb - Martin
* Primer design
-- Documents / E.coli pumps
-- Vectors we could use:
prk404 y7o prk415 resistant to tratracicline, maybe 4 - 9 copies.
pbbr1mcs5 resistant kentamicine up to 10 copies.
puc up to 20 copies resistant to ampicillin.
pjet, resistant to ampicillin up to 600 copies.
REMEMBER not to combine the vector with the same replication origin.
Plbb: clorma resistant, up to 12 copies with LacI in cis (it can be controlled with IPTG)
• Choosing the most suitable gene and designing primers.
Tm = (2 (A + T)) + (4 * (C + G))
• Choose the single cut site!
• Compatibility of restriction sites.
• Read more to start writing.
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