Difference between revisions of "Part:BBa K190019:Experience"

(→‎User Reviews)
(→‎User Reviews)
 
(4 intermediate revisions by 2 users not shown)
Line 28: Line 28:
 
|width='60%' valign='top'|
 
|width='60%' valign='top'|
  
−
We found the sequence of this part to be correct as listed and modified it by the addition of a microcompartment targeting domain and a HA epitope tag (as <partinfo>K562018</partinfo>). This will allow better monitoring of production of the protein. We did not get the chance to test arsenic resistance in a host cell producing this part.
+
We found the sequence of this part to be correct as listed and modified it by the addition of a microcompartment targeting domain and an HA epitope tag (as <partinfo>K562018</partinfo>). This will allow better monitoring of production of the protein. Indeed, we could detect production of this fusion in <i>E. coli</i> using an anti-HA monoclonal antibody. We did not get the chance to test arsenic resistance in a host cell producing this part.
 
|}
 
|}
 
;
 
;
 
<!-- End of the user review template -->
 
<!-- End of the user review template -->
 
<!-- DON'T DELETE --><partinfo>BBa_K190019 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K190019 EndReviews</partinfo>
 +
 +
<!-- DON'T DELETE --><partinfo>BBa_K190019 StartReviews</partinfo>
 +
{|width='80%' style='border:1px solid gray'
 +
|-
 +
|width='10%'|
 +
<partinfo>BBa_K190019 AddReview 2</partinfo>
 +
<I>iGEM Tokyo_NokoGen 2011</I>
 +
|width='60%' valign='top'|
 +
 +
We fused fMT (K190019) to PduP1-18 tag protein (BBa_K519014), that is known to be encapsulated into PduBMC (propanediol-utilization bacterial micro compartment). We observed its growth curve at OD595 in different Cd(II) concentrations, and found that E. coli expressing PduP1~18-fMT resisted medium of higher Cd(II) concentration than the E. coli that does not express fMT. We found that fMT still functions as a metallothionein even when fused to a tag protein, and that it is probably binding to Cd(II). http://2011.igem.org/Team:Tokyo-NoKoGen/metallothionein
 +
|}
 +
;
 +
<!-- End of the user review template -->

Latest revision as of 08:35, 5 October 2011

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_K190019

fMT can be used to sequester metal ions like arsenite (As-III) or arsenate (As-V), this can be used to give the bacterium a higher resistance against arsenic. But the protein can also be used as a delay system together with a metal sensitive promoter (as BBa_K190023), as it sequesters the arsenic as soon as it enters the cell. Upon overexpression of the protein it would take some time before it is saturated and free arsenic can reach the promoter regulator protein (like ArsR). Alternatively the protein can be used to use bacteria for biomining of arsenic, as the presence of rare metals will also become less prominent, like the fossil oil problem the earth just has a maximum amount of some compounds.

User Reviews

UNIQ33841f3c401ee3a7-partinfo-00000001-QINU

••

iGEM Groningen 2009

The capacity of fMT 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 fMT (induced by lactose).

•••

iGEM Dundee 2011

We found the sequence of this part to be correct as listed and modified it by the addition of a microcompartment targeting domain and an HA epitope tag (as BBa_K562018). This will allow better monitoring of production of the protein. Indeed, we could detect production of this fusion in E. coli using an anti-HA monoclonal antibody. We did not get the chance to test arsenic resistance in a host cell producing this part.

UNIQ33841f3c401ee3a7-partinfo-00000005-QINU

UNIQ33841f3c401ee3a7-partinfo-00000006-QINU

••

iGEM Tokyo_NokoGen 2011

We fused fMT (K190019) to PduP1-18 tag protein (BBa_K519014), that is known to be encapsulated into PduBMC (propanediol-utilization bacterial micro compartment). We observed its growth curve at OD595 in different Cd(II) concentrations, and found that E. coli expressing PduP1~18-fMT resisted medium of higher Cd(II) concentration than the E. coli that does not express fMT. We found that fMT still functions as a metallothionein even when fused to a tag protein, and that it is probably binding to Cd(II). http://2011.igem.org/Team:Tokyo-NoKoGen/metallothionein