Creating an E. Coli Strain to Produce Antivenom

Background

Each year snakes envenomate 421,000 people, 20,000 of whom die. These injuries are especially concentrated in developing countries, where snake bites are an occupational hazard. (Kasturiratne et al. 2008). The negative impact of this could be alleviated by the creation and production of a low-cost, human-compatible universal antivenom. Lethal Toxin Neutralizing Factor, henceforth LTNF, is a substance that has been isolated from opossum (Didelphis virginiana) serum, liquid component of blood. LTNF can neutralize nearly all venoms, by a mechanism not yet understood, including those never before encountered by the opossum (Menchaca & Perez 1981, Shier 2008). The active site of LTNF has been isolated into a 15 amino acid-long synthetic oligopeptide designated LT-15 (Lipps 2008 A, Lipps 2008 C). LT-15 also neutralizes venom of many organisms, including snakes, scorpions, jellyfish and bacteria (Lipps 1999, Lipps 2000, Lipps 2003, Lipps 2008 B). Our solution is to create a universal antivenom is modify a strain of Escherichia coli to produce LT-15 (Lipps & Lipps 2005). We will insert the LT-15 gene into a plasmid with a promoter, then transform E. coli with the plasmid and grow the recombinant strain. (Cawood 2013, Cohen et al. 1973, Huang et al. 2012, Lipps 2002 B, Lodish et al. 2000, Muyrersa et al. 2001). These E. coli can then be grown and harvested industrially for LT-15.

This is a novel project because this would be the first universal antivenom and would be among the first synthetic antivenoms. Currently, antivenoms are produced individually for each venom that might need to be neutralized (Domont et al. 1991). LT-15, on the other hand, has been shown to neutralize a multitude of toxins and could be used universall...

... middle of paper ...

.... 623.

Muyrersa, J., Stewart, F. & Zhanga, Y. 2001, "Techniques: Recombinogenic engineering–new options for cloning and manipulating DNA", Trends in Biochemical science, vol. 26, no. 5, pp. 325-331.

Plasmid 44364: pUC-Leum, 2011. Available from: . [6 November 2013].

Shier, W. 2008, “Introduction to the Special Issue on Lethal Toxin Neutralizing Factor", Toxin Reviews, vol. 27, pp. 79-80.

Stocker, J.F. & Traynor, J.R. 1986, "The actions of various venoms on Escherichia coli", Journal of Applied Bacteriology, vol. 61, pp. 383-388.

Xiaoli Dong, Paul Stothard, Ian J. Forsythe, and David S. Wishart "PlasMapper: a web server for drawing and auto-annotating plasmid maps" Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W660-4.

Kapper, J.B. Nataro, J.P. & Mobley, H.L.T. 2004, "PATHOGENIC ESCHERICHIA COLI", Nature Reviews, vol. 2, pp. 123-140.