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Food and Feed Safety and Nutritional Assessment of MON 88017 Corn

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and and regime. AG Bayer property protectionpublishing of contents its parties.data therefore and/oror affiliates. property intellectualthird may its as the of and is regulatory owner. a document any such its reproductiondocument Food and Feed Safety and Nutrit owner this of under of rightsthe this Conclusion Based on Data and Information Evaluated According to FDA’s Policy documentand/or rights to of fall use of the This may distribution,and rights owner subject be the violate copy of and may documentpublication, It exploitation this any permission on Foods from New Plant Varieties prohibited the commercial be ional Assessment of MON 88017 Corn Furthermore,any Consequently,without March 30, 2004 Monsanto # 04-CR-109F FDA BNF 97 Prepared by: Contributors : Submitted by: Monsanto Company 800 NorthMonsanto Lindbergh Company Blvd. St. Louis, MO 63167 # 04-CR-109F Page 1 of 223 and and regime. AG Bayer property protectionpublishing of contents its parties.data therefore and/oror affiliates. property intellectualthird may its as the of and is regulatory owner. a document any such its reproductiondocument owner this of PART I:…………………………………………………………………………… 2 under of this rightsthe Table of Contents…………………………………………………………………. 2 documentand/or of rights to of fall use List of Tables……………………………………………………………………... 6 the List of Figures…………………………………………………………………….. 8 This may distribution,and rights owner Certification………………………………………………………………………. 10 subject Release of Information……………………………………………………………. 11 the be violate Abbreviations and Definitions……………………………………………………. 12 copy of and Narrative Summary……………………………………………………………….. 16 may documentpublication, TABLE OF CONTENTS It exploitation this PART II: Synopsis of consultation summary……………………………………..PART 20 I: any Section 1. Name and address of the submitter…………………………….……... 20 permission prohibited Section 2. The subject of this summary and the plant species from which they the be commercial Section 3. Distinctive designations given to the subject of this summary………. 20 Section 4. Identity and sources of the genetic material introduced into Furthermore,any Consequently,without Section 5. The intendedwere tech derived…………………………………………………….…….. Section 6. The application and uses of MON 88017……………………………. 22 Section 7. The applications for wh MON 88017…………………………………………………………... PART III: Status of submissions to other regulatory agencies…………………... 25 Section 1. Status of submission to USDA-APHIS……………………………….. 25 Section 2. Status of submission to EPA………………………………………….. 25 Section 3. Status of submissions to foreign governments………………………... 26 PART IV: Development of MON 88017………………………………………… 27 Section 1. History and biology of corn…………………………………………... 27 nical effect of MON 88017……………………….. 21 Page Section 2. Characterization1.1. Scientific of thename v and taxonom 1.2. Growth and reproductive characterstics of corn……………………..ich MON 88017 is not29 suitable……….…….. 24 1.3. History of corn development………………………………………… 29 2.1. The 2.1.a. The Arabidopsis thaliana EPSPS transit peptide (CTP2)……….. 32 2.1.b. 2.2. The Section 3. Characterization of the cp4 epsps gene and CP4 EPSPS protein………………………... 31 2.3. T-DNA borders……………………………………………….……... 33 2.4. Genetic elementscp4 epsps outside regulatory of the T-DNA sequences…………………………………. border…………………….. 33 32 cry3Bb1 gene and Cry3Bb1 protein……………………………. 32 3.1. Molecular analysis ic classification of corn……………….. 27 20 3.1.a. Insert and copy number…………………………………………..ector used in transformation………………... 44 31 Monsanto Company3.1.b. cp4 epsps cassette intactness…………………………………….. 46 20 – introducedintroduced DNA…..…………………………... genetic material………………….. 40 40 # 04-CR-109F Page 2 of 223 Table of contents (cont.) Page PART IV. Section 3. (cont.) 3.1.c. p-ract + ract1 intron probe……………………………………….. 46 3.1.d. ctp2 + cp4 epsps probe…………………………………………... 48 3.1.e. NOS 3’ probe……………………………………………………. 48 3.1.f. cry3Bb1 cassette intactness……………………………………… 51 3.1.g. p-e35S probe…………………………………………………….. 51 3.1.h. wtCAB leader + ract1 intron probe……………………….…….. 51 3.1.i. cry3Bb1 probe…………………………………………….…….. 52 3.1.j. tahsp17 3’ probe…………………………………………………. 52 3.2. Analysis for backbone fragments……………………………………. 57 3.3. Insert stability in multiple breeding generations……………………. 59 3.4. Organization of the insert in MON 88017…………………………... 62 3.5. Inheritance of corn rootworm protection and glyphosate toleranceand and regime. traits in MON 88017……………………………….………………..AG 65 3.6. Summary of the molecular characterization of MON 88017………... 67 Section 4. Other data and information about developmentBayer property of MONprotectionpublishing 88017……... 67 of contents its parties.data therefore PART V: Presence of genes that encode resistance to antibiotics…………….….and/oror 68 affiliates. property intellectualthird may its as PART VI: Characterization of thethe prot ofeins introducedand into MON 88017…….…. 69 is regulatory owner. a document Section 1. Identity and function ofany the suchCP4 EPSPS and Cry3Bb1its proteins reproductiondocument present in MON 88017………………………………………………...owner this of 69 under of 1.1. Characterization of therights CP4the EPSPS protein producedthis in documentand/or of rights MON 88017………………………………………………….……...to of fall use 69 the This may distribution,and 1.1.a. Western blot analysisrights to confirmowner the identity of the CP4 EPSPS protein…………………………………………………………...subject 70 be the violate 1.1.b. Molecular weightcopy and purityof of the CP4 EPSPS protein………... 70 and 1.1.c. MALDI-TOFmay document MSpublication, tryptic map mass analysis……………………. 73 It exploitation 1.1.d. N-terminalthis sequence any analysis……………………………………. 73 1.1.e. Functional activity………………………………………………..permission 75 prohibited 1.1.f. Glycosylation analysis……………………………………………the be 75 1.1.g. Conclusionscommercial for characterization of the CP4 EPSPS protein Furthermore,any producedConsequently,without in MON 88017……………………………………….. 77 1.2. Characterization of the Cry3Bb1 protein produced in MON 88017… 77 1.2.a. Western blot analysis to confirm the identity of the Cry3Bb1 protein…………………………………………………………... 78 1.2.b. Molecular weight and purity of the Cry3Bb1 protein……….…… 79 1.2.c. MALDI-TOF MS tryptic map mass analysis…………………….. 82 1.2.d. Functional activity of the Cry3Bb1 protein……………………… 83 1.2.e. Glycosylation analysis of the Cry3Bb1 protein………………….. 83 1.2.f. Conclusions of the characterization of the Cry3Bb1 protein produced in MON 88017…………………………………………. 83 Section 2. Levels of the CP4 EPSPS and Cry3Bb1 proteins in MON 88017……. 86 Monsanto Company # 04-CR-109F Page 3 of 223 and and regime. AG Bayer property protectionpublishing of contents its parties.data therefore and/oror affiliates. property intellectualthird may its as the of and Page is regulatory owner. a document Section 3. Estimate of dietary exposure………………………………………….. 91 any such its reproductiondocument owner this of under of rightsthe this and/or 3.1. Estimated dietary exposure to the CP4 EPSPS protein……………… 91 document to of fall of rights Section 4. Assessment of the potential use 3.2. Estimated dietary exposure to the Cry3Bb1 protein………………… 92 the This may distribution,and 3.3. Margins of exposure…………………………………………………. 92 rights owner subject be the violate Table of contents (cont.) copy of 4.1. AssessmentCry3Bb1 proteins of the potential produced in MON 88017………………………….. and may documentpublication, It exploitation this 4.1.a. Sourceprotein………………………………………………………………. of the CP4 EPSPS protein………………………………... 95 any 4.1.b. Proportion of the total permission prohibited 4.1.c. Bioinformatics analyses of sequence similarity of the CP4 EPSPS the be commercial 4.2. 4.1.d.Assessment Stability of ofthe the potential CP4 EPSPS protei protein produced in MON 88017 to allergens and toxins………… Furthermore,any Consequently,without for allergenicity of 4.2.a. Sourceprotein………………………………………………………………. of the Cry3Bb1 protein……………………………….….. 100 4.2.b. Proportion of the total protein – Cry3Bb1 ………………………. 100 4.2.c. Bioinformatics analyses of sequence similarity of the Cry3Bb1 for allergenicity of the CP4 EPSPS Section 5. Safety assessment of 4.3. 4.2.d.Discussion Stability and of conclusions…………………………………………. the Cry3Bb1 protein in simulated digestiveprotein fluids.. – CP4 106 EPSPS…………………….101 95 protein produced in MON 88017 to allergens and toxins………… 5.1. Safety assessment of the CP4 EPSPS protein……………………….. 106 the CP4 EPSPS and 5.1.a. Structural similarity of th for allergenicity of the Cry3Bb1 5.1.b. Acute oral toxicity study 5.1.c. Safetyand of toxins………………………………………………………….the donor organism: n in simulated digestive fluids.. 96 5.1.d. Similarity of CP4 EPSPS to EPSPSs derived from food sources 5.2. 5.1.e.Safety Presence assessment of CP4 of the EPSPS Cry3Bb1 protein protein………………………….. in food theand CP4 feed EPSPS crops…………. and Cry3Bb1 109 108 proteins………… 106 with a long history of safe consumption………………………….. 5.2.a. Structural similarity of the Cry3Bb1 protein to known proteins 93 5.2.b. Acute oral toxicity study 95 5.2.c. Safetyand of toxins………………………………………………………….the donor organism: e CP4 EPSPS protein to known proteins 5.3. 5.2.d.Conclusions………………………………………………………….. Human and animal safety of 112 with the CP4 EPSPS protein………….. 107 96 Agrobacterium sp. strain CP4……. 107 100 Monsanto Company with the Cry3Bb1 protein……………... 109 101 Bacillus thuringiensis B.t. Cry proteins…………………… 112 106 108 ……………. 111 # 04-CR-109F 109 Page 4 of 223 and and regime. AG Bayer property protectionpublishing of contents its parties.data therefore and/oror affiliates.
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