Part:BBa_K190028:Experience

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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.

UNIQc515d5a31c77c9bb-partinfo-00000005-QINU UNIQc515d5a31c77c9bb-partinfo-00000006-QINU