i c r o g u a r d s OVERVIEW The m i c r o g u a - PowerPoint PPT Presentation

LGE UNICAMP The m i c r o g u a r d s

OVERVIEW The m i c r o g u a r d BIOFUELS s

OVERVIEW The m i c r o g u a r d ETHANOL s

OVERVIEW The m i c • Ideal conditions r o g Costs-benefits u a r d s

OVERVIEW The m i c r o g u a r d s

OVERVIEW The Land use in Brazil m i c Total land area (851 Mha, 100%) r o g Agricultural properties area (355 Mha, 42%) u Cropland area (76.7 Mha, 9%) a Sugarcane cropland for energy-use area r (3.6 Mha, 0.5%) d Source: www.bioetanoldecana.org - Adapted s

OVERVIEW The Raw material utilized to m produce ethanol i c Avoided Energy Raw Material r emissions Produced/Utilized o Sugarcane 9.3 89% g Corn 0.6 – 2.0 -30% to 38% u Wheat 0.97 – 1.11 19% to 47% a Beet 0.97 – 1.11 35% to 56% r Cassava 1.6 – 1.7 63% d Source: www.bioetanoldecana.org s

OVERVIEW The Global ethanol production m i c r o g u a r d Source:Statistical Review of world energy 2009 - Adapted s

OVERVIEW Production and costs of The m ethanol in Brazil i c r o g u a r d s Source: D.L. Gazzoni, Adapted

OVERVIEW The m i ETHANOL c r o g A lot can still be done! u a r d s

OVERVIEW The m i c r o g u a r d s

OVERVIEW Biofuels The m i Good substitutes for c oil-based fuels r o g u Solve the environmental a problems r d s

OVERVIEW Biofuels versus Food The m i c r Biofuels do not compete with o food and land use g u a r d s

OVERVIEW Scientific research The m i c Enable appropriate land use r o g u Should continue - obtain more a efficient biofuels using genetic r engineering d s

OVERVIEW The m ETHANOL i c r o g u a Costs r Project d Land use s

OVERVIEW The How is ETHANOL produced in Brazil? m i c r o g u a r d s

OVERVIEW The How is ETHANOL produced in Brazil? m i c r o g u a r d s

OVERVIEW The How is ETHANOL produced in Brazil? m i c r o g CONTAMINANTS u a r d s

OVERVIEW The iGEM project - CONTAMINATION m i YEAST BACTERIA c r o g u a r d Insulin, yogurt s Ethanol

… The m i Prepare c r yourselves to o g get into the u a microguards´ r d lives s

The m i c r o g u a r d s

The m i c r o g u a r d s

The m i c r o g u a r d s

The m i c r o g u a r d s

The Yeastguard OVERVIEW The m i RECOGNITION c r SYSTEM o g u KILLING a SYSTEM r d s

Recognition system The First Mechanism m (increased lactate sensibility) i c r o g Second Mechanism u (recognizing lactate on the interior of the cell) a r Biobrick submitted! Biobrick submitted! d s

The Yeastguard OVERVIEW The m i RECOGNITION c r SYSTEM o g u KILLING a SYSTEM r d s

killing system The m Biobrick submitted! i c r o g u a r d s

Yeastguard OVERVIEW The m i RECOGNITION c r SYSTEM o g u KILLING a SYSTEM r d s

THE YeastguARd The results m i c Successful assembled biobrick r  BBa_K284002: JEN1 Promoter o  BBa_K284003: Partial DLD promoter g u  BBa_K284023: EYFP regulated by ADH1 promoter (characterization device) a  BBa_K284001: Lysozyme from Gallus gallus r  BBa_K284016: Lysozyme constitutive expression (characterization device) d  BBa_K284017: Lysozyme under control of DLD promoter (characterization device) s

The m i c r o g u a r d s

The coliguard OVERVIEW The m Recognition i system c r o Differentiation g system u a r Killing system d s

Recognition system The m Briefly, i c If contaminants release AI2 If contaminants conjugate with our themselves: coliguard: r o 1. . 1. Induces Py skip steps! g 2. Releases AI2 2. . u 3. Activate Differentiation and 3. Activate Differentiation and a Killing mechanism Killing mechanism r d s

The coliguard OVERVIEW The m Recognition i system c r o Differentiation g system u a r Killing system d s

Differentiation system The m Worker Cells Killer Cells i c r o g u a Amount Amount r d s Basal proportion !

Differentiation system The m i Absence of contaminants: c r o g u a r d s

Differentiation system The m Presence of contaminants: i c r o g u a r d s

Differentiation system The m Presence of contaminants: i c r o g u a r d s

Differentiation system The m i Elimination of contaminants: c r o g u a r d s

Differentiation system The m i Absence of contaminants: c r o g u a r d s

Differentiation system The m i c The slippage mechanism controlling the basal proportions r o g (AGTC) 10 u a r Worker lineage characteristics! d s

Differentiation system The m i c The slippage mechanism controlling the basal proportions r o g (AGTC) 10 (AGTC) 9 u a r Killer lineage characteristics! d s

Differentiation system The m i Integration between the mechanisms of slippage and Cre-Recombinase c r Slippage error doesn’t occurs (most cases): o Conjugation Inhibition System g u a r d Cell Cycle! s Worker lineage

Differentiation system The m Integration between the mechanisms of i slippage and Cre-Recombinase c Slippage error occurs (few cases): r o (AGTC) 9 g u a r d s Killer lineage

Differentiation system The m Enhanced Amount of Killers cells induced by i contaminants c r o g u a r d s

The coliguard OVERVIEW The m Recognition i system c r o Differentiation g system u a r Killing system d s

Killing system The m i The Kamikaze System c r o g The Colicin System u a CeaB lethal gene (colicin E2) into F plasmid r CeiB antidote gene into genomic DNA d s

The coliguard OVERVIEW The m Recognition i system c r o Differentiation g system u a r Killing system d s

THE COLIGUARD The REsults m i c Successful assembled biobrick r  BBa_K284008: Py promoter + RFP device (characterization device) o  BBa_K284031: Cre-Recombinase without ATG start codon g u  BBa_K284022: T4 endolysin under control of T7 promoter (characterization device) a Characterized parts r  BBa_K284022 d s

The coliguard results The m Characterizing BBa_K284022 i  Transformations into E. coli strain C43 (T7 promoter is induced by IPTG) c  Grown inocula were diluted to starter OD=0,2 r  Once OD=0,8 was reached  induction with IPTG o  Incubation for 4 hours at 37⁰C g u a r d s

The coliguard results The m Characterizing BBa_K284022 i c r o g u a r d s

The coliguard results The m Characterizing BBa_K284022 i Plated each culture into solid LB-AMP media c r o g u a r d s

The coliguard results The m Characterizing BBa_K284022 i SDS-PAGE c r o g u a r d s

The Summary m i Accomplishments c  2 contamination control models designed, one for prokaryotes and one for r eukaryotes o  30 new biobrick parts and devices designed g  9 new biobrick parts and devices constructed and submitted to the registry u  4 new biobrick parts or devices were tested a  1 new biobrick device worked as expected (BBa_K284022) r  An existing Biobrick part was characterized (BBa_K112806) d  A new approach to an issue of Human Practice in synthetic biology as it relates to s our project was outlined and detailed

This work could not be done without the sponsor of The m i c r o g u a r d s

The Special Acknowledgements: m Prof. Dr. Fernando Costa. Dean, UNICAMP . i Prof. Dr. Luís Cortez. CORI, UNICAMP . c Prof. Dr. Fábio Papes. Dept. Genética, Evolução e Bioagentes, UNICAMP . r Prof. Dr. Paulo Arruda. Dept. Genética, Evolução e Bioagentes, UNICAMP. o Prof. Dr. Gonçalo A. G. Pereira. Dept. Genética, Evolução e Bioagentes, UNICAMP. g Support: u a r d s

The m i c r thank you!!!! o g u a r d s