IGEM:MIT/2008/Brainstorming

This is the new brainstorming page for our iGEM project. The previous brainstorming page is at IGEM:MIT/2008/Brainstorming Old

---

Expression of a p1025-containing peptide in E. coli

References

PMID 9920267
A synthetic peptide adhesion epitope as a novel antimicrobial agent.

Materials

• inducible promoters
  

(The T7 system has been recommended by protein biochemists for over-expression inside the cell. Should see green cell pellets if GFP is included. Freezing and thawing can disrupt cells followed by extraction)

Adhesion of Streptococcus mutans to hydroxyapatite (HA)

Reference

PMID 9062560
Inhibition of Streptococcus mutans adsorption to hydroxyapatite by low-molecular-weight chitosans.

The materials and methods section describes an indirect plating assay to quantify binding of S. mutans to HA. Results in Table 1 indicates that this method can be used to quantify inhibition of binding by a competitor molecule. In their study, the molecules are sugar polymers.

Materials

• HA beads (from Fluka, 80-200 micro-meter grain size)
• saliva as source of the glyco-protein (one vendor is found through Google)
• S. mutans and handling protocol (Chia is contacting Daniel Smith and Martin Taubman of the Forsyth Institute to acquire a suitable strain)

Secretion of a p1025-containing peptide from Lactobacillus bulgaricus/lactis

References

PMID 12788739 (a secretion signal sequence described; can use the promoter, RBS and secretion signal for this protein for p1025)
Determination of the domain of the Lactobacillus delbrueckii subsp. bulgaricus cell surface proteinase PrtB involved in attachment to the cell wall after heterologous expression of the prtB gene in Lactococcus lactis.

PMID 11772607
Electrotransformation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis with various plasmids.

Materials

• Lactobacillus bulgaricus or lactis (check with Tom Knight first. Otherwise request from another lab or purchase from ATCC)
• growth medium: Ligia recommends purchasing the pre-mix powder for the MRS medium for technical simplicity
• Gas Pak to create anaerobic environment. It looks like (Email bio related mailing lists to see if we can borrow one.)
• Expression vector: BBa_I742103 in registry is a shuttle vector for E. coli and Lactobacillus. Let's look up this vector to see whether people have used it for protein expression. Alternatively we can get some of the plasmids described in PMID 11772607.

Fundrai$ing

Drew has said that he can get in touch with some MIT alumni and the MIT development office. We can send them the brochure and a letter to explain our goal, project and budget.