IGEM:MIT/2008/Notebook/Yogurt

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==Description==
==Description==
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We seek to express the 20aa peptide P1050 in Lactobacillus bulgaricus/lactis.
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*Our goal is to engineer a common yogurt bacteria, Lactobacillus bulgaricus, so that it will express the 20aa peptide p1025. A clinical study [http://www.ncbi.nlm.nih.gov/pubmed/9920267?dopt=Abstract (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.
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We will express P1050 in E. coli as an intermediate step to evaluate protein secretion and efficacy.
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** 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.
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** Stage 2: Binding Assay - see if the p1025 produced by E.coli inhibits binding of ''S. mutants'' to hydroxyapatite (HA) beads.
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** Stage 3: Expression of p1025 in Lactobacillus.
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** Stage 4: Make yogurt with modified Lactobacillus and test for inhibition of ''S. mutants'' binding.
==Notes==
==Notes==
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* Group meetings every Friday at 4pm in 68-574
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* Group meetings every Friday at 4pm in 68-474 (except 7/11 and 8/1 in 68-674)
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* Journal club meetings as needed every Friday at 11am in 68-474
==Information==
==Information==
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* [[../Yogurt/Brainstorming |Brainstorming]]
* [[../Yogurt/Notes |Daily Notebook]]
* [[../Yogurt/Notes |Daily Notebook]]
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* [[../Yogurt/Label |Labeling Information]]
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* [[../Yogurt/sequences |Primers/Plasmids/PCR sequences]]
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* [[../Yogurt/lbulgaricus |''Lactobacillus delbrueckii'' subsp. ''bulgaricus'' Information & Protocols]]
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* [[../Yogurt/toothbindingassay | Tooth binding assay protocol]]
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* [[../Yogurt/Vocabulary List |Vocabulary List]]
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* [[../Yogurt/teamleader |Team Leader Responsibilities]]
==Useful Links==
==Useful Links==
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*[http://slam.bs.jhmi.edu/gd/ |Peptide Sequence Manipulations]
*[http://slam.bs.jhmi.edu/gd/ |Peptide Sequence Manipulations]
*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]
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*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 9062560<br style="clear:both;"/> Inhibition of Streptococcus mutans adsorption to hydroxyapatite by low-molecular-weight chitosans.<br style="clear:both;"/>
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*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;"/>
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*PMID 11772607<br style="clear:both;"/> Electrotransformation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis with various plasmids.

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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 68-474 (except 7/11 and 8/1 in 68-674)
  • Journal club meetings as needed every Friday at 11am in 68-474

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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