IGEM:MIT/2008/Notebook/Yogurt

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==Description/Abstract==
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==Description==
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* Place short description of project or notes regarding this project
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* Lorem ipsum dolor sit amet, consectetuer adipiscing elit. Donec porta commodo tellus. Nam a est eget libero mollis tincidunt. Aliquam purus. Quisque nulla ligula, facilisis in, pulvinar sed, molestie a, quam. Vestibulum at pede. In in sem eget odio eleifend placerat. Phasellus ultricies felis quis sapien. Etiam molestie volutpat quam. Praesent pulvinar scelerisque mi. Nam mi urna, fringilla eu, mattis sed, venenatis id, nunc. Maecenas velit eros, congue ut, placerat in, ornare vel, sem. Aenean porta enim sit amet felis gravida posuere. Phasellus faucibus nibh et orci.
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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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** 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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* Place some notes here for visitors
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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
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==Information==
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* [[../Yogurt/Brainstorming |Brainstorming]]
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* [[../Yogurt/Notes |Daily Notebook]]
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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]]
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==Useful Links==
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*[http://www.biology.utah.edu/jorgensen/wayned/ape/ |DNA Plasmid Editor]
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*[http://slam.bs.jhmi.edu/gd/ |Peptide Sequence Manipulations]
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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]
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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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* Example: Next team meeting on Tuesday, June 18th! Be there!
 
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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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