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2 11/075593 Al (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date , , / r » - 23 June 2011 (23.06.2011) 2 11/075593 Al (51) International Patent Classification: James Calvin [US/US]; 3123 Evergreen Road, Ames, C12N 15/82 (2006.01) C07K 14/325 (2006.01) Iowa 50014 (US). SCELONGE, Christopher Jay A01H 1/02 (2006.01) A01H 5/00 (2006.01) [US/US]; 1406 N.W. Cedarwood Drive, Ankeny, Iowa 50021 (US). VILLANELO, M. Alejandra Pascual [CL/ (21) International Application Number: US]; 1910 West Street, Granger, Iowa 50109 (US). PCT/US20 10/060841 YOUNG, Gregory James [US/US]; 2128 The Highway, (22) International Filing Date: Wilmington, Delaware 19810 (US). YOUNG, Joshua K. 16 December 2010 (16.12.2010) [US/US]; 5981 Somerset Place, Johnston, Iowa 5013 1 (US). ZHONG, Cathy Xiaoyan [CN/US]; 2922 Jaffe (25) Filing Language: English Road, Wilmington, Delaware 19809 (US). (26) Publication Langi English (74) Agent: MOXON, Claire L.; Pioneer Hi-Bred Interna (30) Priority Data: tional, Inc., 7250 N.W. 62nd Avenue, Johnston, Iowa 61/287,492 17 December 2009 (17.12.2009) US 5013 1-0552 (US). 61/413,659 15 November 2010 (15.1 1.2010) US (81) Designated States (unless otherwise indicated, for every (71) Applicants (for all designated States except US): PIO¬ kind of national protection available): AE, AG, AL, AM, NEER HI-BRED INTERNATIONAL, INC. [US/US]; AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, 7100 N.W. 62nd Avenue, Johnston, Iowa 5013 1-1014 CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, (US). E. I. DUPONT DE NEMOURS & COMPANY DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, [US/US]; 1007 Market Street, Wilmington, Delaware HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, 19898 (US). KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, (72) Inventors; and NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD, (75) Inventors/ Applicants (for US only): DIEHN, Scott [US/ SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, US]; 4720 Tamara Lane, West Des Moines, Iowa 50265 TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (US). LU, Albert L. [CA/US]; 14 Charles Pointe, Newark, Delaware 19702 (US). NOWATZKI, Timothy (84) Designated States (unless otherwise indicated, for every M. [US/US]; 2402 Maple Street, Granger, Iowa 50109 kind of regional protection available): ARIPO (BW, GH, (US). NUBEL, Douglas Stuart [US/US]; 12428 Sunset GM, KE, LR, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, Terrace, Clive, Iowa 50325 (US). REGISTER, III, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, [Continued on next page] (54) Title: MAIZE EVENT DP-040416-8 AND METHODS FOR DETECTION THEREOF 40416 maize (57) Abstract: The invention provides DNA compositions that relate to transgenic insect resistant maize plants. Also provided are assays for detecting the presence of the maize DP-040416-8 event based on the DNA sequence of the recombinant construct in- serted into the maize genome and the DNA sequences flanking the insertion site. Kits and conditions useful in conducting the as- says are provided. EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, Published: LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, — with international search report (Art. 21(3)) SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, ML, MR, NE, SN, TD, TG). — with sequence listing part of description (Rule 5.2(a)) MAIZE EVENT DP-04041 6-8 AND METHODS FOR DETECTION THEREOF FIELD OF INVENTION Embodiments of the present invention relate to the field of plant molecular biology, specifically embodiment of the invention relate to DNA constructs for conferring insect resistance to a plant. Embodiments of the invention more specifically relate to insect resistant corn plant event DP-04041 6-8 and to assays for detecting the presence of corn event DP-04041 6-8 in a sample and compositions thereof. BACKGROUND OF INVENTION An embodiment of this invention relates to the insect resistant corn (Zea mays) plant DP-04041 6-8, also referred to as "maize line DP-04041 6-8," "maize event DP-04041 6-8," and "4041 6 maize," and to the DNA plant expression construct of corn plant DP-04041 6-8 and the detection of the transgene/flanking insertion region in corn plant DP-04041 6-8 and progeny thereof. Corn is an important crop and is a primary food source in many areas of the world. Damage caused by insect pests is a major factor in the loss of the world's corn crops, despite the use of protective measures such as chemical pesticides. In view of this, insect resistance has been genetically engineered into crops such as corn in order to control insect damage and to reduce the need for traditional chemical pesticides. One group of genes which have been utilized for the production of transgenic insect resistant crops is the delta-endotoxin group from Bacillus thuringiensis (Bt). Delta-endotoxins have been successfully expressed in crop plants such as cotton, potatoes, rice, sunflower, as well as corn, and have proven to provide excellent control over insect pests. (Perlak, F.J etal. ( 1 990) Bio/Technology 8:939-943; Perlak, F.J. et al. ( 1 993) Plant Mol. Biol. 22:31 3-321 ; Fujimoto, H . et al. ( 1 993) Bio/Technology 11: 15 1- 1 155; Tu et al. (2000) Nature Biotechnology 18:1 10 1- 1 104; PCT publication WO 01/1 3731 ; and Bing, J.W. et al. (2000) Efficacy of Cry1 F Transgenic Maize, 14th Biennial International Plant Resistance to Insects Workshop, Fort Collins, CO). The expression of foreign genes in plants is known to be influenced by their location in the plant genome, perhaps due to chromatin structure (e.g., heterochromatin) or the proximity of transcriptional regulatory elements (e.g., enhancers) close to the integration site (Weising et al. ( 1 988) Ann. Rev. Genet. 22:421 -477). At the same time the presence of the transgene at different locations in the genome will influence the overall phenotype of the plant in different ways. For this reason, it is often necessary to screen a large number of events in order to identify an event characterized by optimal expression of an introduced gene of interest. For example, it has been observed in plants and in other organisms that there may be a wide variation in levels of expression of an introduced gene among events. There may also be differences in spatial or temporal patterns of expression, for example, differences in the relative expression of a transgene in various plant tissues, that may not correspond to the patterns expected from transcriptional regulatory elements present in the introduced gene construct. For this reason, it is common to produce hundreds to thousands of different events and screen those events for a single event that has desired transgene expression levels and patterns for commercial purposes. An event that has desired levels or patterns of transgene expression is useful for introgressing the transgene into other genetic backgrounds by sexual outcrossing using conventional breeding methods. Progeny of such crosses maintain the transgene expression characteristics of the original transformant. This strategy is used to ensure reliable gene expression in a number of varieties that are well adapted to local growing conditions. It would be advantageous to be able to detect the presence of a particular event in order to determine whether progeny of a sexual cross contain a transgene of interest. In addition, a method for detecting a particular event would be helpful for complying with regulations requiring the pre-market approval and labeling of foods derived from recombinant crop plants, for example, or for use in environmental monitoring, monitoring traits in crops in the field, or monitoring products derived from a crop harvest, as well as for use in ensuring compliance of parties subject to regulatory or contractual terms. It is possible to detect the presence of a transgene by any nucleic acid detection method known in the art including, but not limited to, the polymerase chain reaction (PCR) or DNA hybridization using nucleic acid probes. These detection methods generally focus on frequently used genetic elements, such as promoters, terminators, marker genes, etc., because for many DNA constructs, the coding region is interchangeable. As a result, such methods may not be useful for discriminating between different events, particularly those produced using the same DNA construct or very similar constructs unless the DNA sequence of the flanking DNA adjacent to the inserted heterologous DNA is known. For example, an event- specific PCR assay is described in U.S. Patent No. 6,395,485 for the detection of elite event GAT-ZM1 . Accordingly, it would be desirable to have a simple and discriminative method for the identification of event DP-04041 6-8. SUMMARY OF INVENTION Embodiments of this invention relate to methods for producing and selecting an insect resistant monocot crop plant. More specifically, a DNA construct is provided that when expressed in plant cells and plants confers resistance to insects. According to one aspect of the invention, a DNA construct, capable of introduction into and replication in a host cell, is provided that when expressed in plant cells and plants confers insect resistance to the plant cells and plants.
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