IGEM:MIT/2008/Notebook/Yogurt

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

• 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.