IGEM:MIT/2008/Notebook/Yogurt

< IGEM:MIT | 2008 | Notebook(Difference between revisions)

Current revision

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.

|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.

@@ Line 23: / Line 23: @@
 ==Notes==
-* Group meetings every Friday at 4pm in 68-574
+* 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==
@@ Line 33: / Line 34: @@
 * [[../Yogurt/sequences |Primers/Plasmids/PCR sequences]]
-* [[../Yogurt/Label |Labeling system]]
+* [[../Yogurt/lbulgaricus |''Lactobacillus delbrueckii'' subsp. ''bulgaricus'' Information & Protocols]]
+* [[../Yogurt/toothbindingassay | Tooth binding assay protocol]]
+* [[../Yogurt/Vocabulary List |Vocabulary List]]
+* [[../Yogurt/teamleader |Team Leader Responsibilities]]
 ==Useful Links==
@@ Line 44: / Line 51: @@
 *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;"/>
 *PMID 11772607<br style="clear:both;"/> Electrotransformation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis with various plasmids.