The document proposes research to investigate whether the bacterium Pseudomonas stutzeri can synthesize the neurotoxin saxitoxin. It describes using bioinformatics to search for cyanobacterial saxitoxin genes in P. stutzeri strains. Preliminary results found some genes with strong similarity, suggesting horizontal gene transfer, while other genes had weaker matches. The research aims to amplify and sequence these genes from a P. stutzeri marine strain to explore its potential to produce saxitoxin.

1. Building Genetic Foundations in the saxitoxin Production of Pseudomonas stutzeri A Research Proposal Gabrielle Roberts

15. Repolarization Rest State Passer, M.W. & Smith, R. E. (2004). Psychology the Science of Mind and Behavior: Second Edition. New York: McGraw Hill. Depolarization Repolarization

17. a-subunit Ion conducting pore The pore loop between the S5 and S6 segments 1. Na+ enter 2. Is the site where saxitoxin binds “ Inner ring pore loop”

18. SAXTITOXIN CHEMICAL STRUCTURE The guanidine, hydroxyl and carbamoyl groups of saxitoxin bind to the voltage gated sodium channel’s inner ring pore loop

20. Transfer of guanidino group to intermediate Claisen condensation of arginine and acetate Kellman, R., Mihali, T. K., Jeon, Y. J., Pickford, R., Pomati, F., & Neilan, B. A. (2008). Biosynthetic Intermediate Analysis and Functional Homology Reveal a Saxitoxin Gene Cluster in Cyanobacteria. Applied and Environmental Microbiology , 74, No.13, 4044-4053. carbamoylation hydroxylation

21. SAXTITOXIN MOLECULAR SYNTHESIS SxtG Guanidino transfer SxtB SxtD Cyclization Methylation Epoxidation SxtS SxtS opening SxtU Reduction Carbamoylation SxtH/T SxtW SxtV Hydroxylation SxtI Claisen condensation: acetate to arginine

30. Preliminary Results

34. Preliminary Results Good E-values Carbamoyl Transferase 7e-88 Short-chain dehydrogenase 3e-40 8-amino-7-oxononanoate 9e-44 Closest Blast Match in P. stutzeri A1501 STX Enzyme in C.raciborskii T3 sxtI sxtU sxtA

35. Carbamoyl transferase E-value: 7e-88 Exceptional E-value is evidence of horizontal gene transfer between Cylindrospermopsis raciborskii T3 and Pseudomonas stutzeri A1501 Pseudomonas stutzeri A1501

37. Preliminary Results Iron sulfur Protein 0.001 Sterol desaturase 0.037 Tyosine decarboxylase putative 0.063 Closest Blast Match in P. stutzeri A1501 STX Enzyme in C.raciborskii T3 sxtW sxtD sxtJ sxtB Hypothetical protein PST_2386 0.14 Chemotaxis Protein CheV sxtG 1.9 E-value