IGEM:MIT/2008/Notebook/Yogurt

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*Primer Analyzer Tool (check temperature & GC content) [http://www.idtdna.com/analyzer/Applications/OligoAnalyzer/Default.aspx | One] [http://www.idtdna.com/analyzer/Applications/OligoAnalyzer/Default.aspx | Two]
*Primer Analyzer Tool (check temperature & GC content) [http://www.idtdna.com/analyzer/Applications/OligoAnalyzer/Default.aspx | One] [http://www.idtdna.com/analyzer/Applications/OligoAnalyzer/Default.aspx | Two]
*PMID 9920267<br style="clear:both;"/> A synthetic peptide adhesion epitope as a novel antimicrobial agent.<br style="clear:both;"/>
*PMID 9920267<br style="clear:both;"/> A synthetic peptide adhesion epitope as a novel antimicrobial agent.<br style="clear:both;"/>
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*PMID 12536251<br style="clear:both;"/> Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems.<br style="clear:both;"/>
 
*PMID 9062560<br style="clear:both;"/> Inhibition of Streptococcus mutans adsorption to hydroxyapatite by low-molecular-weight chitosans.<br style="clear:both;"/>
*PMID 9062560<br style="clear:both;"/> Inhibition of Streptococcus mutans adsorption to hydroxyapatite by low-molecular-weight chitosans.<br style="clear:both;"/>
*PMID 12788739<br style="clear:both;"/> (a secretion signal sequence described; can use the promoter, RBS and secretion signal for this protein for p1025).<br style="clear:both;"/>
*PMID 12788739<br style="clear:both;"/> (a secretion signal sequence described; can use the promoter, RBS and secretion signal for this protein for p1025).<br style="clear:both;"/>

Revision as of 09:11, 17 June 2008

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Description

  • Our goal is to engineer a common yogurt bacteria, Lactobacillus bulgaricus, so that it will express the 20aa peptide p1025. A clinical study (Kelly CG et al.; Nature Biotechnol. 1999) reports that p1025 is good for your teeth. p1025 reduces oral colonization of Streptococcus mutans, a tooth-decaying bacterium.
    • Stage 1: Construction of p1025 fusion peptide and expression of gene in E. coli. This is an intermediate step to evaluate gene function and protein secretion/efficacy.
    • Stage 2: Binding Assay - see if the p1025 produced by E.coli inhibits binding of S. mutants to hydroxyapatite (HA) beads.
    • Stage 3: Expression of p1025 in Lactobacillus.
    • Stage 4: Make yogurt with modified Lactobacillus and test for inhibition of S. mutants binding.

Notes

  • Group meetings every Friday at 4pm in 68-574

Information

Useful Links

  • |DNA Plasmid Editor
  • |Peptide Sequence Manipulations
  • Primer Analyzer Tool (check temperature & GC content) | One | Two
  • PMID 9920267
    A synthetic peptide adhesion epitope as a novel antimicrobial agent.
  • PMID 9062560
    Inhibition of Streptococcus mutans adsorption to hydroxyapatite by low-molecular-weight chitosans.
  • PMID 12788739
    (a secretion signal sequence described; can use the promoter, RBS and secretion signal for this protein for p1025).
  • PMID 11772607
    Electrotransformation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis with various plasmids.




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