Difference between revisions of "Part:BBa K190028:Experience"

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We did not have a lot of trouble with doing a pcr of GlpF to get it out of the genome or to get the EcoR1 and Pst1 site out. We used it in testing the import of arsenic by doing a accumulation assay and analyse it with ICP-MS. Protocols can be found [http://2009.igem.org/Team:Groningen/Protocols here]
 
We did not have a lot of trouble with doing a pcr of GlpF to get it out of the genome or to get the EcoR1 and Pst1 site out. We used it in testing the import of arsenic by doing a accumulation assay and analyse it with ICP-MS. Protocols can be found [http://2009.igem.org/Team:Groningen/Protocols here]
 
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The capacity of GlpF to enhance the arsenite accumulation in ''E. coli'' was tested using an arsenite uptake [http://2009.igem.org/Team:Groningen/Protocols assay]. But unfortunately the arsenic concentration determined by ICP-MS was not reproducible and did not show enhanced arsenite accumulation in ''E. coli'' carrying GlpF (induced by lactose).
 
The capacity of GlpF to enhance the arsenite accumulation in ''E. coli'' was tested using an arsenite uptake [http://2009.igem.org/Team:Groningen/Protocols assay]. But unfortunately the arsenic concentration determined by ICP-MS was not reproducible and did not show enhanced arsenite accumulation in ''E. coli'' carrying GlpF (induced by lactose).
 
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The capacity of GlpF to transport arsenite over the membrane in ''E. coli'' was tested using this [http://2009.igem.org/Team:Groningen/Protocols#Death_assay assay]. The [http://2009.igem.org/Team:Groningen/Project/Transport#Results  results] show working GlpF constructs.
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Latest revision as of 21:21, 21 October 2009

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_K190028

This part has been used to import As(III) into the E.coli cells. It has been used in combination with metal accumulation proteins to make a accumulation device. The two devices are BBa_K190038, a combination of GlpF with the accumulatin protein [http://2009.igem.org/Team:Groningen/Project/Accumulation#Metals fMT] and BBa_K190037, a combination of GlpF with the fusion protein of MBP with [http://2009.igem.org/Team:Groningen/Project/Accumulation#Metals ArsR] (an accumulation protein).

User Reviews

•••••

iGEM_Groningen2009

We did not have a lot of trouble with doing a pcr of GlpF to get it out of the genome or to get the EcoR1 and Pst1 site out. We used it in testing the import of arsenic by doing a accumulation assay and analyse it with ICP-MS. Protocols can be found [http://2009.igem.org/Team:Groningen/Protocols here]


••

iGEM_Groningen2009

The capacity of GlpF to enhance the arsenite accumulation in E. coli was tested using an arsenite uptake [http://2009.igem.org/Team:Groningen/Protocols assay]. But unfortunately the arsenic concentration determined by ICP-MS was not reproducible and did not show enhanced arsenite accumulation in E. coli carrying GlpF (induced by lactose).


••••

iGEM_Groningen2009

The capacity of GlpF to transport arsenite over the membrane in E. coli was tested using this [http://2009.igem.org/Team:Groningen/Protocols#Death_assay assay]. The [http://2009.igem.org/Team:Groningen/Project/Transport#Results results] show working GlpF constructs.

UNIQf984f44038ceffe8-partinfo-00000005-QINU UNIQf984f44038ceffe8-partinfo-00000006-QINU