Team:University College London/Module 5

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=Module 5: Salt Tolerance=
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== Description ==
== Description ==
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A core module for our project is enabling E.coli to '''tolerate''' the '''salt content''' of the ocean; without this ability E.coli '''could not survive''' in a marine environment. This system is not required for other bacteria we will be transforming, '''Roseobacter denitrificans''' and '''Oceanibulbus indolifex''', which as native '''marine''' bacteria already have salt tolerance. Rather, because of the widespread use of E.coli as a chassis for Synthetic Biology this Module is being developed to demonstrate that E.coli too could have the capacity to carry the plastic formation or degradation system.  
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A core module for our project enables ''E. coli'' to '''survive''' the '''salt concentration''' of the ocean - without this ability the application of ''E. coli'' to a marine environment would not be possible. This system would not be required for the other bacteria strains we will be transforming, '''''Roseobacter denitrificans''''' and '''''Oceanibulbus indolifex''''', which as native '''marine''' bacteria already have genes conferring salt tolerance. Due to the widespread use of ''E. coli'' as a chassis for synthetic biology, this module is being developed to demonstrate that ''E. coli'' could be applied to marine environments.  
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We propose to confer salt tolerance on E.coli by linking the salt tolerance gene encoding the protein irrE BBa_K729001 to the constitutive promoter BBa_J23119. The choice of a constitutive promoter is intuitive; E.coli must sustain expression of salt tolerant proteins in order to survive.  
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The protein '''IrrE''' originates from '''''Deinococcus radiodurans''''', where it confers resistance to radiation. When transformed into ''E. coli'' however, it protects against salt, oxidative and thermal shock. IrrE appears to function as a '''global regulator''' of stress factor genes. So far it has been demonstrated to upregulate transcription of '''recA''' and '''pprA''' – genes which encode '''Recombinase A''' and '''Radiation Inducible Protein'''. With respect to salt tolerance, IrrE upregulates the production of several '''stress responsive''' proteins, protein '''kinases''', '''metabolic''' proteins, and '''detoxification''' proteins. It also downregulates '''glycerol degradation'''. With this global regulatory effect, ''E. coli'' becomes more salt tolerant.
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The protein''' irrE''' is found naturally in '''Deinococcus radiodurans''', where it confers resistance to radiation. When transformed into E.coli however, it protected against salt, oxidative and thermal shock. irrE appears to work by regulation of a number of genes. So far it has been demonstrated to upregulate transcription of '''recA''' and '''ppr'''A – genes which encode '''Recombinase A''' and '''Radiation Inducible Protein'''. This may be involved in promoting DNA repair and protecting against oxidative damage when transformed into bacteria, but the exact mechanism of salt tolerance is not clear. We will specifically investigate the ability of irrE to confer salinity tolerance in E. coli, as previously demonstrated by Pan et al. 2009.
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We propose to confer salt tolerance on ''E. coli'' by linking the salt tolerance gene encoding the protein IrrE (BBa_K729001) to a constitutive promoter (BBa_J23119). The choice of a constitutive promoter is intuitive; ''E. coli'' must sustain expression of salt tolerant proteins in order to survive.  
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Latest revision as of 00:24, 27 September 2012

Description | Design | Construction | Characterisation | Modelling | Results | Conclusions

Description

A core module for our project enables E. coli to survive the salt concentration of the ocean - without this ability the application of E. coli to a marine environment would not be possible. This system would not be required for the other bacteria strains we will be transforming, Roseobacter denitrificans and Oceanibulbus indolifex, which as native marine bacteria already have genes conferring salt tolerance. Due to the widespread use of E. coli as a chassis for synthetic biology, this module is being developed to demonstrate that E. coli could be applied to marine environments.

The protein IrrE originates from Deinococcus radiodurans, where it confers resistance to radiation. When transformed into E. coli however, it protects against salt, oxidative and thermal shock. IrrE appears to function as a global regulator of stress factor genes. So far it has been demonstrated to upregulate transcription of recA and pprA – genes which encode Recombinase A and Radiation Inducible Protein. With respect to salt tolerance, IrrE upregulates the production of several stress responsive proteins, protein kinases, metabolic proteins, and detoxification proteins. It also downregulates glycerol degradation. With this global regulatory effect, E. coli becomes more salt tolerant.

We propose to confer salt tolerance on E. coli by linking the salt tolerance gene encoding the protein IrrE (BBa_K729001) to a constitutive promoter (BBa_J23119). The choice of a constitutive promoter is intuitive; E. coli must sustain expression of salt tolerant proteins in order to survive.