IGEM:MIT/2008/Notebook/Yogurt

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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.

Information

 * Brainstorming


 * Daily Notebook


 * Primers/Plasmids/PCR sequences


 * Lactobacillus delbrueckii subsp. bulgaricus Information & Protocols


 * Tooth binding assay protocol


 * Vocabulary List


 * Team Leader Responsibilities

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.