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Revision as of 03:41, 27 October 2012 (view source) Manugimenez (Talk | contribs) (→Data Page) ← Older edit		Latest revision as of 03:54, 27 October 2012 (view source) Manugimenez (Talk | contribs) (→Data Page)
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	[[File:Crossfeeding-v03-small.png | 700px]]		[[File:Crossfeeding-v03-small.png | 700px]]
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		+	In order for the cross-feeding scheme to work we need each strain to export the amino acid they produce (either Histidine or Tryptophan). To achieve this we created a devices design to secrete to the medium an His (or Trp) rich peptides.
		+
		+	{| style="width: 100%"
		+	| align="center" | [[File:Crossfeeding-device-design_v01.jpg]]
		+	|-
		+	| align="center" | '''Basic DNA structure of the devices with their constitutive parts'''
		+	|}
		+
		+
		+	* '''Kozak''' consensus sequence for initiation of translation.
		+	* '''Signal peptide''' that targets the product of the gene for secretion.
		+	* '''Trojan peptide''' to increase internalization in target cell.
		+	* '''Payload''': this is the exported amino acid rich domain of the protein.
		+
		+	The input of the devices are ''PoPS'' and the output is secreted amino acids, so the devices are ''PoPS -> exported AA'' transducers. In principle any PoPS generating part can be used.
		+
		+
		+	You can read more about the design process [[Team:Buenos_Aires/Results/Bb1 | here]]
	We built 2 '''His-export devices''', and 2 '''Trp-export devices''':		We built 2 '''His-export devices''', and 2 '''Trp-export devices''':

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Latest revision as of 03:54, 27 October 2012

• iGEM BsAs
  • The team
  • Official roster
  • Members
  • Where we come from

  • Safety
  • Key questions

  • Attributions
  • Attributions

  • Sponsors
  • Institutional

  Data Page
• Project
  • Synthetic ecology
  • Overview
  • Motivation
  • Design
  • Possible applications

  • Schemes
  • Crossfeeding
  • Independent Population Control
  • Cross-population control
  • Stochastic State Transitions

  • Modeling
  • Modeling synEcology
  • Advanced modeling

  Data Page
• Results and Devices
  • Strains characterization
  • Strains description
  • Fluorescence screening
  • Screening of strain proportion
  • Auxotrophy confirmation
  • Coculture in liquid medium
  • Revertant control
  • Trp basal production
  • Growth dependence

  • BBs design
  • Trp/His export devices
  • Backup devices
  • Preparing for sending

  • Testing
  • BioBricks testing
  • SynEco testing

  Data Page
• Human practices
  • Garage Lab
  • Spreading the word
  • Solving local problems

  • EMBO
  • Seeding SynBio in Latin America

  Data Page

Data Page

In order for the cross-feeding scheme to work we need each strain to export the amino acid they produce (either Histidine or Tryptophan). To achieve this we created a devices design to secrete to the medium an His (or Trp) rich peptides.

Basic DNA structure of the devices with their constitutive parts

• Kozak consensus sequence for initiation of translation.
• Signal peptide that targets the product of the gene for secretion.
• Trojan peptide to increase internalization in target cell.
• Payload: this is the exported amino acid rich domain of the protein.

The input of the devices are PoPS and the output is secreted amino acids, so the devices are PoPS -> exported AA transducers. In principle any PoPS generating part can be used.

You can read more about the design process here

We built 2 His-export devices, and 2 Trp-export devices:

• His-export I (<partinfo>BBa_K792009</partinfo>)
• Trp export I (<partinfo>BBa_K792010</partinfo>)
• His-export II (<partinfo>BBa_K792011</partinfo>)
• Trp export II (<partinfo>BBa_K792012</partinfo>)

To achive this, we had to create several new basic biobricks:

• Kozak sequence from yeast α-factor mating pheromone (MFα1) (<partinfo>BBa_K792001</partinfo>)
• Secretion tag from yeast α-factor mating pheromone (MFα1) (<partinfo>BBa_K792002</partinfo>)
• HIV TAT penetratin (<partinfo>BBa_K792003</partinfo>)
• Polyarginine trojan peptide (<partinfo>BBa_K792004</partinfo>)
• PolyHa, a Histidine rich peptide (His-Tag) (<partinfo>BBa_K792005</partinfo>)
• TrpZipper2, a Tryptophan rich peptide water soluble and monomeric (<partinfo>BBa_K792006</partinfo>)
• PolyHb, a stable Histidine rich peptide designed by us (<partinfo>BBa_K792007</partinfo>)
• PolyWb, a stable Tryptophan rich peptide designed by us(<partinfo>BBa_K792008</partinfo>)