Team:Newcastle/Metalsensing

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|'''[https://2009.igem.org/Team:Newcastle/Labwork/27_August_2009#Metal_Sensor_Team 27th August 2009]'''
|'''[https://2009.igem.org/Team:Newcastle/Labwork/27_August_2009#Metal_Sensor_Team 27th August 2009]'''
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|Digested ''pSB1A2'' (containing ''BBa_J33206'' BioBrick)
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|Digested ''pSB1A2'' (containing ''BBa_J33206'' BioBrick), ran DNA though gel and excised band.
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|'''[https://2009.igem.org/Team:Newcastle/Labwork/28_August_2009#Metal_Sensor_Team] 28th August 2009'''
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|Cleaned gel band using Gel extraction kit
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Revision as of 21:13, 21 October 2009


Metal Sensing

Introduction

If our project is to process cadmium and not other metals, we need to genetically engineer Bacillus subtilis to carry out a set of cellular processes based on the action of metal sensors. These metal sensors will detect cadmium through a system known as AND Gating.

There are two metal sensing repressors, which are known to respond to cadmium: arsR and czrA. By placing binding sites to these two metal sensing repressors next to each other in a promoter region, the gene regulated by that promoter will be synthesized only when a combination of metals that bind to both sensors are present; this is a combinatorial approach for gene expression regulation.

Modelling

BioBrick constructs

Lab Work Strategies

Other Presentations and Diagrams

Newcastle Metalsensor2100.gif

Lab Work done

Summary of Lab Sessions for Cadmium Sensing
Date
Description
18th August 2009 Transformed DH5-alpha E. coli cells with BBa_J33206 from the Spring Distribution
19th August 2009 Inoculated 3 tubes of LB with 3 colonies of potential transformant E.coli cells
20th August 2009 Conduct mini-preps on the three overnight-grown cultures of potential BBa_J33206 transformants and digested them with restriction enzymes.
21st August 2009 Analysed digested BBa_J33206 mini-prep DNA by DNA gel electrophoresis and prepared midi-preps also.
25th August 2009 Concentrated (and ethanol precipitated) the BBa_J33206 Midi-prep sample.
26th August 2009 Digested BBa_J33206 midi-prep DNA with EcoRI and PstI and analysed through DNA gel electrophoresis - digest reaction not successful (a second attempt at digests needed)
27th August 2009 Digested pSB1A2 (containing BBa_J33206 BioBrick), ran DNA though gel and excised band.
[1] 28th August 2009 Cleaned gel band using Gel extraction kit



News

Events

Social Net

  • Newcastle iGEM Twitter
  • [http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
  • [http://www.youtube.com/user/newcastle2009igem Newcastle Youtube Channel]
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