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=== Stage 1 === | === Stage 1 === | ||
| - | *Construction of p1025 fusion peptide and expression of gene in E. coli. This is an intermediate step to evaluate gene | + | *Construction of p1025 fusion peptide with appropriate markers and expression of gene in E. coli. This is an intermediate step to evaluate gene construction and protein efficacy |
===Stage 2=== | ===Stage 2=== | ||
| - | *Binding Assay - | + | *Binding Assay - To reproduce the results of the Nature paper and test if the p1025 construct made in stage 1 inhibits binding of Streptococcus mutans to hydroxyapatite beads as well as the original p1025 peptide. |
===Stage 3=== | ===Stage 3=== | ||
| - | *Expression of p1025 in Lactobacillus | + | *Expression of p1025 in Lactobacillus bulgaricus |
===Stage 4=== | ===Stage 4=== | ||
*Make yogurt with modified Lactobacillus and test for inhibition of ''S. mutants'' binding | *Make yogurt with modified Lactobacillus and test for inhibition of ''S. mutants'' binding | ||
Revision as of 15:32, 16 October 2008
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Overall project
A clinical study (Kelly CG et al.; Nature Biotechnol. 1999) reports that a short synthetic peptide (20 amino acids long) called p1025 can reduce oral colonization of a major tooth-decaying bacterium Streptococcus mutans and thus help to maintain oral health. The peptide does so by competitively inhibit binding of S. mutans to a glycoprotein from saliva on tooth surface. The peptide is considered advantageous because it selectively prevents S. mutans colonization without removing beneficial bacteria that are also present in the mouth.
Our research goal is to engineer one of the common yogurt bacteria, Lactobacillus bulgaricus, so that it secrets p1025 in yogurt. Consumption of the yogurt after a meal could reduce tooth decay. We are not going to market this yogurt. (Neither will any of us taste this yogurt! There’s no telling what residual chemicals could be left over from lab work) However, our project will demonstrate a bio-engineering approach to increase the value of common probiotics.
Project Details
Stage 1
- Construction of p1025 fusion peptide with appropriate markers and expression of gene in E. coli. This is an intermediate step to evaluate gene construction and protein efficacy
Stage 2
- Binding Assay - To reproduce the results of the Nature paper and test if the p1025 construct made in stage 1 inhibits binding of Streptococcus mutans to hydroxyapatite beads as well as the original p1025 peptide.
Stage 3
- Expression of p1025 in Lactobacillus bulgaricus
Stage 4
- Make yogurt with modified Lactobacillus and test for inhibition of S. mutants binding
The Experiments
- For daily information about wet-lab experiments performed, please follow directions in the "Procedural Notebook section" of [[1]]
- For any further details regarding experiments, please refer to the appropriate sub-sections of [[2]]
Results
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