IGEM:Harvard/2007/Bacterial Targeting

Bacterial Targeting (LppOmpA)

Goals

• To make bacteria adhere to targets with a high degree of specificity.
• To make bacteria transduce a signal upon adhesion to their targets.

Planned Work

6/27/07

Transform strains of our controls - strep and his
Prepare them in liquid culture

6/28/07

Innoculate and Induce controls Run them on the MACS system to see if the assay works.

Readings

1. Wang JY and Chao YP. Immobilization of cells with surface-displayed chitin-binding domain. Appl Environ Microbiol. 2006 Jan;72(1):927-31. DOI:10.1128/AEM.72.1.927-931.2006 |
2. Georgiou G, Stephens DL, Stathopoulos C, Poetschke HL, Mendenhall J, and Earhart CF. Display of beta-lactamase on the Escherichia coli surface: outer membrane phenotypes conferred by Lpp'-OmpA'-beta-lactamase fusions. Protein Eng. 1996 Feb;9(2):239-47. DOI:10.1093/protein/9.2.239 |
3. Stathopoulos C, Georgiou G, and Earhart CF. Characterization of Escherichia coli expressing an Lpp'OmpA(46-159)-PhoA fusion protein localized in the outer membrane. Appl Microbiol Biotechnol. 1996 Mar;45(1-2):112-9. DOI:10.1007/s002530050657 |
4. Henning U, Cole ST, Bremer E, Hindennach I, and Schaller H. Gene fusions using the ompA gene coding for a major outer-membrane protein of Escherichia coli K12. Eur J Biochem. 1983 Nov 2;136(2):233-40. DOI:10.1111/j.1432-1033.1983.tb07732.x |
5. Francisco JA, Earhart CF, and Georgiou G. Transport and anchoring of beta-lactamase to the external surface of Escherichia coli. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2713-7. DOI:10.1073/pnas.89.7.2713 |
6. Chen W and Georgiou G. Cell-Surface display of heterologous proteins: From high-throughput screening to environmental applications. Biotechnol Bioeng. 2002 Sep 5;79(5):496-503. DOI:10.1002/bit.10407 |
7. Earhart CF. Use of an Lpp-OmpA fusion vehicle for bacterial surface display. Methods Enzymol. 2000;326:506-16. DOI:10.1016/s0076-6879(00)26072-2 |
8. Dane KY, Chan LA, Rice JJ, and Daugherty PS. Isolation of cell specific peptide ligands using fluorescent bacterial display libraries. J Immunol Methods. 2006 Feb 20;309(1-2):120-9. DOI:10.1016/j.jim.2005.11.021 |
9. Nakajima H, Shimbara N, Shimonishi Y, Mimori T, Niwa S, and Saya H. Expression of random peptide fused to invasin on bacterial cell surface for selection of cell-targeting peptides. Gene. 2000 Dec 30;260(1-2):121-31. DOI:10.1016/s0378-1119(00)00461-3 |