I hereby certify that this paper or fee is being deposited with the United States Postal Service "EXPRESS MAIL POST OFFICE T0 Signature
`
`NUMBER EL 831818998 US
`
`DATE OF DEPOSIT February 14, 2005
`
`EXPRESS MAIL MAILING LABEL
`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`TRADEMARK TRIAL AND APPEAL BOARD
`
`Registration No.: 2,833,109
`Mark: GA21
`
`Registered: April 13, 2004
`
`SYNGENTA CROP PROTECTION, INC.,
`
`Petitioner,
`
`
`
`
`V.
`
`Cancellation No. 92043653
`
`MONSANTO TECHNOLOGY LLC,
`
`Registrant.
`
`MOTION TO SUSPEND PROCEEDINGS
`
`PENDING OUTCOME OF FEDERAL COURT LITIGATION
`
`Pursuant to 37 C.F.R. § 2.117(a) and Trademark Trial and Appeal Board Manual of
`
`Procedure (“TBMP”) § 510.02, Registrant Monsanto Technology LLC (“Registrant”) hereby
`
`moves to suspend this cancellation proceeding pending resolution of two ongoing federal court
`
`cases. In support of this motion to suspend, Registrant states as follows:
`
`1.
`
`Petitioner Syngenta Crop Protection, Inc. (“Petitioner”) initiated this cancellation
`
`proceeding on September 1, 2004, seeking to cancel Registrant’s United States Trademark
`
`Registration No. 2,833,109 (“the ’ 109 Registration”) for the mark GA21. The ’ 109 Registration
`
`covers a “plant derived protein which confers in-plant tolerance to glyphosate herbicides.”
`
`DM_US\8156720.v1
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`IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
`
`02-14-2005
`U.S. Pawn! & TMOfcITM Mai! Rap! Dt. #64
`
`

`

`
`
`The Delaware Federal Patent Case
`
`2.
`
`Registrant is currently involved in a case pending in federal court in Delaware
`
`against Syngenta Seeds, Inc. and Syngenta Biotechnology, Inc. These two Syngenta companies
`
`are, like Petitioner herein, business divisions of Syngenta AG. This federal court case is
`
`encaptioned Monsanto Company and Monsanto Technology LLC v. Syngenta Seeds, Inc., and
`
`Syngenta Biotechnology, Inc., Civ. No. 04-305 SLR (D. Del.) (hereinafter the “Delaware case”).
`
`3.
`
`In the Delaware case, Registrant asserts that Petitioner’s sister divisions are
`
`infringing U.S. Patent No. 4,940,835, entitled “Glyphosate Resistant Plants” (the “ ’835 patent”).
`
`(See Delaware Amended Complaint 111] 6-7, 9-11.)’ The ’835 patent concerns a plant gene that
`
`confers tolerance in the plant to glyphosate herbicides. (See id.)
`
`4.
`
`In the Delaware case, Registrant seeks to enjoin Petitioner’s sister divisions and
`
`all those in privity with them from infringing the ’835 patent by making, using, selling or
`
`offering to sell products containing the gene taught by the ’835 patent that confers tolerance to
`
`glyphosate herbicides. (See id. at p. 4.) The defendants in the Delaware case have answered.
`
`(See Exh. B to Casagrande Decl.)
`
`The Illinois Federal Patent Case
`
`5.
`
`Petitioner’s sister divisions, Syngenta Seeds, Inc. and Syngenta Biotechnology,
`
`Inc. are also defendants in another pending patent case encaptioned DEKALB Genetics
`
`Corporation v. Syngenta Seeds, Inc, and Syngenta Biotechnology, Inc., Civ. No. 04C50323 (N .D.
`
`Ill.) (hereinafter the “Illinois case”).
`
`The Delaware Amended Complaint is Exhibit A to the Declaration of Thomas L. Casagrande, Exh. 1
`
`1 h
`
`ereto.)
`
`DM_US\8 1 56720.vl
`
`

`

`
`
`6.
`
`In the Illinois case, Petitioner’s sister divisions are alleged to infringe U.S. Patents
`
`Nos. 5,538,880, entitled “Method For Preparing Fertile Transgenic Corn Plants” (the “ ’880
`
`patent”) and 6,013,863, entitled “Fertile Transgenic Corn Plants” (the “ ’863 patent”).
`
`7.
`
`In the Illinois case, Petitioner’s sister divisions are alleged to infringe the ’880
`
`patent by “making corn containing genes that confer resistance to the herbicide glyphosate and
`
`importing into the United States, or offering to sell, selling, or using within the United States,
`
`corn containing genes that confer resistance to the herbicide glyphosate made by the process of
`
`one or more claims 1-9 of the ’88O Patent.” (See Amended Complaint 111] 13-14, 16, 11-l2.)2
`
`8.
`
`In the Illinois case, Petitioner’s sister divisions are also alleged to infringe the
`
`’863 patent by “making corn containing genes that confer resistance to the herbicide glyphosate
`
`and importing into the United States, or offering for sale, selling, or using within the United
`
`States, corn containing genes that confer resistance to the herbicide glyphosate made by the
`
`process of the ’863 Patent.” (See id.)
`
`9.
`
`In the Illinois case, the complaint asks the court to enter an injunction against
`
`Petitioner’s sister divisions and all those in privity with them from infringing claims 1-9 of the
`
`’880 patent and the claims of the ’863 patent. (See id. at p. 7.) Petitioner’s sister divisions have
`
`not yet filed
`
`answer to the pending amended complaint in the Illinois case.
`
`The Delaware and Illinois Cases May Have a Bearing on This Case
`
`10.
`
`37 C.F.R. § 2.117(a) states:
`
`Whenever it shall come to the attention of the Trademark Trial and Appeal
`Board that parties to a pending case are engaged in a civil action which
`may have a bearing on the case, proceedings before the Board may be
`suspended until termination of the civil action.
`
`The Illinois Complaint is Exhibit C to the Casagrande Declaration, Exh. 1.
`
`DM_US\8l56720.v1
`
`3
`
`

`

`See also General Motors Corp. v. Cadillac Club Fashions Inc., 22 USPQ2d 1933, 1936-37
`
`(TTAB 1992); Marie Claire Album S.A. v. Kruger GmbH & Co. KG, 29 USPQ2d 1792, 1794
`
`(TTAB 1993); TBMP § 510.02(a) (“Ordinarily, the Board will suspend proceedings in the case
`
`before it if the final determination of the other proceeding will have a bearing on the issues
`
`' before the Board”) The rationale for this rule is that, “[t]o the extent that a civil action in
`
`Federal district court involves issues in common with those in a proceeding before the Board, the
`
`decision of the Federal district court is often binding upon the Board .
`
`.
`
`. .” TBMP § 510.02(a).
`
`11.
`
`The final determination in the Delaware and Illinois cases will have a bearing on
`
`this cancellation proceeding. Specifically, although the instant Petition is not a model of clarity,
`
`it appears that Petitioner alleges that Registrant’s GA2l mark is “descriptive and generic” for the
`
`plant gene identified in the description of goods and services “which confers in-plant tolerance to
`
`glyphosate herbicides.” (Petition W 5, l 1.)
`
`12.
`
`Petitioner also alleges in this proceeding that Petitioner uses GA2l or a similar
`
`term in Petitioner’s advertising and “on the identical products of Petitioner.” (Id. 1} 7) (emphasis
`
`added). Thus, the primary asserted basis for Petitioner’s cancellation petition appears to be that
`
`it uses GA21 descriptively in connection with the sale of a plant gene “which confers in—plant
`
`tolerance to glyphosate herbicides.”
`
`13.
`
`Indeed, the very reason why Petitioner alleges that it would be damaged by the
`
`continued registration of GA2l is that “[c]ontinued registration of the Trademark will lead the
`
`public to conclude, incorrectly, that Registrant has superior rights to use the Trademark in
`
`conjunction with the sale ofplant traits or proteins for glyphosate tolerance to certain herbicides
`
`and other products, all to the damage of Petitioner .
`
`.
`
`. .” (Id 1112) (emphasis added).
`
`DM_US\8 l 56720.vl
`
`

`

`
`
`14.
`
`If the patentees in the either or both of the Delaware or Illinois cases prevail,
`
`Petitioner’s sister divisions and anyone in privity with them (such as other divisions of the same
`
`parent entity)3 would have no right to make, use, or sell the goods identified in the ’ 109
`
`Registration, i. e., plants containing glyphosate resistant genes.
`
`15.
`
`Only a person “who believes that he or she will be damaged” by a registration has
`
`standing to bring a petition to cancel. 15 U.S.C. § 1064. Facts establishing standing “are part of
`
`a petitioner’s case and must be affirmatively proved.” Lipton Indus., Inc. v. Ralston Purina C0,,
`
`670 F.2d 1024, 1028 (CCPA 1982). Standing to bring an action—like this one—based on
`
`allegations that the mark is merely descriptive of the goods in connection with which it is used,
`
`requires a petitioner to prove that it has a legal right to use the mark to describe its goods. E. g. ,
`
`Jewelers Vigilance Comm. v. Ullenberg Corp., 823 F.2d 490, 493 (Fed. Cir. 1987); Nobelle. com
`
`LLC v. Qwest Communications Int ’I Inc., 66 USPQ2d 1300, 1304 (TTAB 2003). Though this is
`
`not a stringent test, it carmot be met if Petitioner is enjoined by a federal court from making,
`
`using, or selling the very product it says GA21 describes.
`
`16.
`
`It is therefore clear that the Delaware and Illinois cases may indeed have a bearing
`
`on this proceeding:
`
`the outcome of these cases may take away the basis for Petitioner to claim it
`
`is damaged by the continued registration of GA21, thus exposing Petitioner’s lack of standing to
`
`contest the ’ 109 registration. There is no good reason to waste the parties’ time and money, as
`
`well as the Board’s time and resources, to rule on whether “GA21” is descriptive of, or generic
`
`for, a product that Petitioner may well have no legal right to make or sell.
`
`. upon the parties to the
`.
`. is binding .
`.
`See Fed. R. Civ. P. 65(d) (“Every order granting an injunction .
`3
`action, their officers, agents, servants, employees, and attorneys, and upon those persons in active concert and
`participation with them who receive actual notice of the order .
`.
`. .”) (emphasis added); see also Regal Knitwear Co.
`v. NLRB, 324 U.S. 9, 14 (1945) (“a decree of injunction not only binds the parties defendant but also those identified
`with them in interest, in ‘privity’ with them, represented by them or subject to their control”); Walling v. Reuter,
`321 U.S. 671, 674-75 (1944) (an injunction binds any related company under same control as named party);
`
`DM_US\8 l 56720.vl
`
`

`

`WHEREFORE, Registrant prays that this petition to cancel be suspended pending the outcome
`
`of the Delaware and Illinois federal cases.
`
`Dated: February 14, 2005
`
`Fax: (713) 787-1440
`e-mail: casagrandet@howrey.com
`
`Attorneysfor Registrant Monsanto Technology
`LLC
`
`Computer Searching Serv. Corp. v. Ryan, 439 F.2d 6, 9 (2d Cir. 1971) (an injunction would bind the subsidiary of
`the named party under rule 65(d)).
`
`DM_US\8 l 56720.vl
`
`

`

`CERTIFICATE OF SERVICE
`
`I hereby certify that a true and correct copy of the foregoing MOTION TO SUSPEND
`
`PROCEEDINGS PENDING OUTCOME OF FEDERAL COURT LITIGATION has been
`
`served on February 14, 2005 by e-mail and first class mail, postage prepaid, on:
`
`James A. Zellinger, Esq.
`Trademark Counsel
`
`Syngenta Crop Protection, Inc.
`410 Swing Rd.
`Greensboro, N.C. 27409
`
`e-mail: jim.ze11inger@syngenta.com .
`
`DM_US\8 I 56720.vl
`
`

`

`
`
`
`
`Exhibit 1Exhibit 1Exhibit 1
`
`

`

`
`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`
`TRADEMARK TRIAL AND APPEAL BOARD
`
`Registration No.: 2,833,109
`Mark: GA2l
`
`Registered: April 13, 2004
`
`Cancellation No. 92043653
`
`SYNGENTA CROP PROTECTION, INC.,
`
`Petitioner,
`
`V.
`
`MONSANTO TECHNOLOGY LLC,
`
`Registrant.
`
`
`
`DECLARATION OF THOMAS L. CASAGRANDE IN SUPPORT
`
`OF MOTION TO SUSPEND PROCEEDINGS PENDING
`
`OUTCOME OF FEDERAL COURT LITIGATION
`
`I, THOMAS L. CASAGRANDE, pursuant to 28 U.S.C. § 1746, hereby declare:
`
`1.
`
`I am a partner in the law firm of Howrey Simon Arnold & White, LLP, attorneys
`
`of record for Registrant, Monsanto Technology LLC.
`
`I am a member in good standing of the
`
`State Bars of Texas and Connecticut. This declaration is based on my own personal knowledge
`
`2.
`
`My law firm, Howrey Simon Arnold & White LLP, is also counsel of record for
`
`the plaintiffs in the following pending federal court cases: Monsanto Company and Monsanto
`
`Technology LLC v. Syngenta Seeds, Inc., and Syngenta Biotechnology, Inc., Civ. No. 04-305
`
`SLR (D. Del.) (the “Delaware case”); and DEKALB Genetics Corporation v. Syngenta Seeds,
`
`Inc, and Syngenta Biotechnology, Inc., Civ. No. 04C50323 (N.D. I11.) (the “Illinois case”)
`
`3.
`
`Attached hereto as Exhibit A is a true and correct copy of the current complaint in
`
`the Delaware case.
`
`4.
`
`Attached hereto as Exhibit A is a true and correct copy of the current answer in
`
`the Delaware case.
`
`

`

`5.
`
`Attached hereto as Exhibit A is a true and correct copy of the current complaint in
`
`the Illinois case.
`
`Ideclare underpenalty ofperjury that theforegoing is true and«
`
`Dated: February 14, 2005
`
`
`
`
`

`

`
`
`
`
`
`
`
`
`
`
`Exhibit AExhibit AExhibit A
`
`

`

`' ‘T
`
`{_—';
`:;..'
`..
`
`IN THE UNITED STATES DISTRICT COURT
`FOR THE DISTRICT OF DELAWARE
`
`C. A. No. 04-305 SLR
`
`JURY TRIAL DEMANDED
`
`) 3
`
`)
`)
`)
`
`) ) ) )
`
`MONSANTO COMPANY and
`
`MONSANTO TECHNOLOGY LLC,
`
`Plaintiffs,
`
`V-
`
`SYNGENTA SEEDS, INC.; and
`SYNGENTA BIOTECHNOLOGY, INC.
`
`Defendants.
`
`FIRST AMENDED COMPLAINT
`
`Plaintiffs, Monsanto Company and Monsanto Technology LLC (collectively
`
`“Monsanto”), file this first amended complaint against Defendants Syngenta Seeds, Inc.
`
`and Syngenta Biotechnology, Inc. (collectively “Defendants”) prior to any responsive
`
`pleading by Defendants and alleges as follows:
`
`THE PARTIES
`
`1.
`
`Plaintiffs Monsanto‘ Company and Monsanto Technology LLC are both
`
`corporations organized and existing under the laws of the State of Delaware, with their
`
`principal places of business at 800 North Lindbergh Boulevard, St. Louis, Missouri
`
`63167.
`
`2.
`
`On information and belief, Defendant, Syngenta Seeds, Inc. (“Syngenta Seeds”),
`
`is a corporation organized and existing under the laws of the State of Delaware, with its
`
`offices at 7500 Olson Memorial Highway, Golden Valley, Minnesota 55427
`
`3.
`
`On information and belief, Defendant, Syngenta Biotechnology, Inc. (“Syngenta
`
`Biotechnology”), is a corporation organized and existing under the laws of the State of
`
`

`

`
`
`THE INFRINGEMENT
`
`8.
`
`Monsanto realleges and incorporates by reference each of paragraphs 1-7 above
`
`as set forth herein.
`
`9.
`
`Monsanto has been the owner of all right, title and interest to and under United
`
`States Patent No. 4,940,835 entitled “Glyphosate—Resistant Plants.” U.S. Patent
`
`4,940,835 (“the ‘835 Patent”) was duly and legally issued to Monsanto on July 10, 1990.
`
`A copy of the patent is attached as Exhibit 1 to this Complaint.
`
`10.
`
`Defendants do not have any license or other right to practice the claims of U.S.
`
`Patent No. 4,940,835.
`
`11.
`
`Upon information and belief, Defendants have infringed and continue to infiinge
`
`one or more claims of the ‘835 Patent by at least making, using, selling and offering for
`
`sale corn products exhibiting resistance to glyphosate herbicides, and will continue -to do
`
`so unless enjoined by this Court.
`
`12.
`
`Upon information and belief, Defendants have infringed one or more of the
`
`claims of the ‘835 Patent by at least inducing others and contributing to the infringement
`
`by others.
`
`13.
`
`Defendants’ acts of infringement of the ‘835 Patent, upon information and belief,
`
`have been carried out in deliberate and willful disregard of Monsanto’s patent rights.
`
`JURY DEMAND
`
`14.
`
`Pursuant to Rule 38(b), Fed. R. Civ. P., Plaintifls request a trial by jury.
`
`PRAYER FOR RELIEF
`
`WHEREBY PLAINTIFFS PRAY FOR THE FOLLOWING RELIEF:
`
`A.
`
`A judgment that Defendants have infringed the ‘835 Patent;
`
`

`

`
`
`Delaware, with offices located at 3054 Cornwallis Road, Research Triangle Park, North
`
`Carolina 27709-2257.
`
`JURISDICTION AND VENUE
`
`4.
`
`This is an action for patent infringement arising under the Patent Laws of the
`
`United States, Title 35, United States Code § 1 et seq. Subject matter jurisdiction is
`
`proper under 28 U.S.C. §§ 1331 and 1338.
`
`5.
`
`Venue is proper in this Court under 28 U.S.C. §§ 1391 and 1400(b).
`
`BACKGROUND
`
`6.
`
`Plaintiff Monsanto is a leader in the development of crops that have been
`
`genetically engineered to express new traits of value to farmers, such as herbicide
`
`resistance. Products developed by Monsanto include com, soybeans and other crops
`
`containing genes that confer resistance to the herbicide glyphosate. Glyphosate is a
`
`commercial herbicide that kills plants by binding to a critical enzyme in the plant called
`
`“EPSPS.” Monsanto’s glyphosate herbicide is sold under the trademark Roundup® and
`
`the genetically engineered products made by Monsanto that have the glyphosate
`
`resistance trait are sold under the trademark Roundup Ready®. Roundup Ready®
`
`products have been a recognized commercial success. Since the introduction of Roundup
`
`Ready® crops in 1996, farmers have consistently increased the number of acres they
`
`plant in the United States with Roundup Ready® products.
`
`7.
`
`Recognizing the value of Monsanto’s glyphosate resistance technology,
`
`Defendants have attempted to make and use glyphosate resistant corn products and have
`
`conspired with others to make glyphosate resistant corn products in violation of
`
`Monsanto’s patent rights.
`
`

`

`
`
`B.
`
`A judgment that Defendants have willfully and deliberately infringed the ‘835
`
`Patent;
`
`C.
`
`A declaration by the Court that any making, using, selling or offering for sale by
`
`Defendants of any corn products that are within the scope of the ‘835 Patent
`
`would constitute an act of infringement of the ‘835 Patent;
`
`D.
`
`A preliminary and final injunction enjoining Defendants and all those in privy
`
`with it from infringing, from inducing infringement, and from conuibuting to the
`
`infringement of the ‘835 Patent by making, using, selling or offering for sale corn
`
`products that are within the scope of the ‘835 Patent;
`
`E.
`
`An award of compensatory and exemplary damages, but not
`
`less than a
`
`reasonable royalty, resulting from Defendants’ infringement, including allowance
`
`of multiplied damages based on Defendants’ willful and deliberate infringement;
`
`F.
`
`G.
`
`An award of interest, costs, and attorneys’ fees; and
`
`Such other and further relief as this Court shall deem just and proper.
`
`Respectfully submitted,
`
`OF COUNSEL:
`
`POTTER ANDERSON & CORROON LLP
`
`John F. Lynch
`Susan K. Knoll
`
`Steven G. Spears
`HOWREY, SIMON, ARNOLD &
`WHITE, LLP
`750 Bering Drive, #400
`Houston, Texas 77057
`Telephone (713) 787-1400
`
`By:
`
`-2
`Richard L. Horwitz (#2243
`David E. Moore (#3983)
`Hercules Plaza, 6”’ Floor
`1313 N. Market Street
`Wilmington, DE 1980]
`Telephone (302) 984-6000
`
`Dated: June 9, 2004
`
`Attorneys for Plaintiffs
`
`638050
`
`

`

`
`
`
`
`EXHIBIT 1EXHIBIT 1EXHIBIT 1
`
`

`

`
`
`United States Patent [191
`Shah et al.
`Jul. 10, 1990
`[45] Date of Patent:
`
`[11] Patent Number:
`
`4,940,835
`
`[54] GLYPHOSATB-RI-LSETANT PLANTS
`[75]
`Inventors: Dilip M. Shah, Creve Coeur, Stephen
`G. R0361, Chestcrfield; Robert B.
`Horsela; Robert '1'. Finley, both St.
`Louis. 1111 of Mo.
`[73] Assignee: Monsanto Company, St. Louis. Mo.
`[21] Appl. No.: 879,814
`[22]
`1-116.1;
`.1111. 7, 1986
`
`[63]
`
`[51]
`
`Related US. Application Data
`o1'Ser. No. 797.390, Oct. 29, 1985.
`abandzmed, which is a continuation-in-part of Ser. No.
`763.482. Aug. 7. 1935. abandoned.
`Int. c1.= ............3. M1111 1/04; C12N 15/00.
`C12N 5/011. 00711 15/12
`soo/205; 435/177.3;
`[52] US. (:1.
`435/240.4; 435/320; 536/27; 935/35; 935/47;
`935/48; 935/64; 935/67; 8111/1316. 44-;
`Sm/DIG. 43; B00/DIG. 26; 8CD/DIG. 14;
`8(1)/DIG. 17; 800/DIG. 63; Sm/DIG. 27;
`800/1316. 24; 800/230; 8110/0161. 67
`[58] Field of Search ...................... 435/172.3. 68, 317.
`435/240. 240.4, 320; S36/27; 47/58; 300/1
`References Cited
`U.S. PATENT DOCUMENTS
`4.535.060 8/198$ Comai ................_.... 435/317
`5769.061
`9/1988 Co1nai..._..........._..._............ 71/86
`FOREIGN PATENT S
`
`[56]
`
`0115673 8/1984 European Pat. OE. ........... 935/14
`OTHER PUBLICATIONS
`
`Smart et al., J. of 3101 Chem (1935) 260, 30. pp.
`16338-16346.
`Lee, ‘r. '1‘. J. Plant Growth Regal (1932) 137-43.
`Jensen. R. A. P1193161 Plant (1935) 66:164-168.
`Rubin et 31., Plant Physial (1932) 70, 333-339.
`Mousdale et 11.. Planza (1935) 163-241-249.
`’dAmato. -r. A, 6: 31., Pla:1ta(1984) 162:104—108.
`Rothe, G. M., et al. Ham (1933) 157-353-366.
`Singhse: 31.. Arch. qffiioclmn 3 Biaph. (1935) 2432 pp.
`374-3
`.
`Saijo et a1., Agric. Biol. Chem (1979) 43 (7) pp.
`1427-1432
`Schmidt et al., nuns (1933) so, 2632-2636.
`
`Stalker ct 31-. J. Biol. Chem (1985) 260:8 pp. 4724-4728.
`Science (1986) 231: 1360-1361.
`Biotechnology (1984) Nov., pp. 944.
`Cole et 131., Weed Research (1983) 23. 173-183.
`Rubin et al. Plant PI1ys1'ol (1984)7S, pp. 839-945.
`Guilley et al. (1982) “Transcription of CAMV DNA. .
`.” Cell 30: 763-73.
`Mazur et al. (1985) “Cloning Herbicide. . . " World
`Biotcch Rep 2: 97-108.
`Goodman et al. (1987) “Gene Transfer in Crop.
`Science 236: 48-54.
`Amrheim et al. (1983) “Biochemical Basis For.
`FEBS Letters 157: 191-6.
`Naja'geretal. (1934) “Selection & Chaiacterizntion. . ."
`Plant Physiol 76: 571-4.
`Vanden Broeck et al. (1985) "Targeting of. . ." Nature
`313: 358-63.
`Koziel et al. (1984) “A Cntiliflower Mosiac Virus. . ." J.
`Mol. Appl. Genet 2: 549-62.
`Roget: et a1 (1983) Appl. Environ Microbiol 46 : 37-43.
`Comai et al. (1983) Science 221: 370-1.
`Primary ExamIuer—Charles F. Warren
`Asistant Exarniner—David '1'. Fox
`Attorney. Agent. orFrn1—Dennis R. I-Ioemer, Jr.; James
`W. Williams. 11:; Larry R. Swaney
`
`.
`
`."
`
`. ."
`
`ABSTRACT
`[57]
`This invention involves a cloning or expression vector
`comprising 9. gene which codes 5-enoIpyruvylshiki-
`mate-3-phosphate
`synthase
`(EPSPS)
`polypeptide
`w11ich,w11enexprmedinap1antoellconta.insachlo1'o-
`plast transit peptide which allows the polypeptide, or an
`enzymntically nctive portion thereof. to be transported
`from the cytoplasm of the plant cell into a chloroplast in
`thep1antoel1,andconfersasubstantia1degreeofg,1y-
`phosate raistance upon the plant cell and plants regen-
`erated therefrom.
`
`The EPSPS coding sequence may be ligated to a strong
`promoter, such 3 the 35S promoter from cauliflower
`xnosaicyirus, to create 11 chimeric gene. Such genes can
`be inserted into plant transformation vectors. and subse-
`quently introdnced into plant cells Plant cells trans-
`formed using such genes and plants regenerated there-
`from have b shown to exhibit a substantial degree of
`glyphosate resistance.
`
`59C1lnims,8Drnw'lngSheets
`
`1%’:
`fig?-
`12/
`!\‘E§§
`2;
`as
`
`.
`
`IONIIJIJIIAIVO
`
`

`

`
`
`US. Patent
`
`Jul. 10, 1990
`
`Sheet '1 of8
`
`4,940,835
`
`2. CONTACT WITH LOW LEVEL OF GLYPHOSATE
`AND CULTURE SURVIVING CELLS
`
`3. SLOWLY INCREASE LEVEL OF GLYPHOSATE, ISOLATE
`HIGHLY RESISTANT (Gr ) CELLS, MP4 - G
`
`
`
`4. PURIFY EPSPS
`PROTEIN: DETERMINE AMINO
`ACID SEQUENCE
`
`6. CREATION or cDNA
`snow PETUNIA rr.FINA
`
`_
`
`5. CREATE RADIOACTIVE DNA
`PROBES wmcn COULD eucooe
`. e MINO ACID sequence
`
`7. INSERT CDNA INTO
`
`CLONING vecrons
`
`3. use moses TO ISOLATE A CLONING vecron
`HAVING AN eesps GENE m msemen FRAGMENT
`
`.
`
`9. DETERMINE SEQUENCE OF EPSPS GENE, IDENTIFY
`CLEAVAGE SITES THAT ALLOW PROMOTER TO BE
`REMOVED FROM CODING SEQUENCE
`
`10. REPLACE EPSPS PROMOTER WITH STRONGER CaMV 35S .
`PROMOTER, TO CREATE CHIMERIC GENE
`
`11. INSERT CHIMERIC GENE INTO DISARMED TI VECTOR.
`USE VECTOR TO INSERT THE GENE INTO PLANT CELLS
`
`12. GROW TRANSFORMED PLANT CELLS ON GLYPHOSATE
`
`13. REGENERATE AND TEST DIFFERENTIATED PLANTS
`
`‘
`
`I
`
`FIGURE 1
`
`

`

`US. Patent
`
`Jul. 10, 1996
`
`Sheet 2 of 8
`
`4,940,835
`
`\NOS
`
`BACTERIAL
`Spc I Str R
`
`
`
`pBR 322
`
`om.
`
`Psx
`
`ass
`
`puIoN 530
`CaMV
`
`-
`
`"03 3
`
`Bgm
`Pstl
`Eco RI
`
`/
`
`Nos - NPTII - Nos
`(PLANT Kenfi)
`
`“U1
`Egigl.
`I
`T‘ “"55
`’
`
`~
`
`PMO_N 53s
`
`-
`
`Eco RI,
`CAP,
`7‘ “W55
`
`-
`
`.
`
`Eco R!
`Bglll
`s- I——4—————I——I
`5'
`CTP REGION
`N-T
`REGION
`' EPSPS INSERT FROM
`pNIoN 9531
`
`TRUNCATED
`EPSPS
`
`Eco RI
`Eco RI
`|——————o———I
`TRUNCATED
`POLY-A
`UNKER
`EPSPS
`TAIL
`EPSPS INSERT FROM
`pMON 9556
`
`tms tmr tml ocs
`
`DELETE I
`
`
`Kan R
`
`430 Kb
`
`Pstl
`
`Vir
`
`on
`
`One
`
`5°" "1
`
`HELPER PLASMID
`
`Cam I EPSPS I NOS
`
`FIGURE 2
`
`

`

`U.S. Patent
`
`Jul. 10, 1990
`
`Sheet 3 of 8
`
`4,940,835
`
`IGIATTCCCTCAAICTTTACTTICAAGAATGGCACMATTAACAACATGGCACAAGGGAIA 60
`ueunaolnneAsnAsnueuueIncIy1 le
`
`51 CAMCCCITAATCCCAATTCC AATTTCCATIAACCCCAAGTTCCTAAATCTTCAAGTTTT
`GI nThrLeu|snPr oksnsu AsnPheHi sLysPr oGl nVa| ProLys5erSerSerPhe
`
`120
`
`121 CTTGTTTTTGGATCTAAAAAACTGAAMATTCAGCAAATTCTATGTTGGTTTTGAAAAAA 1&0
`Leuval Phelil ysertyslys LeuLysAsnserAI altsnset letuuval LeuLysLys
`
`181GATTCAATTTTTATGCAAAAGTTTTGTTCCTTTAGGATTTCAGCATCAGTGGCTACAGCA 240
`Aspsetll ePhe|ln GI nlys PhacysSuPheAvgIl eser Al aserval Al aThrAl a
`
`241 CAG
`GI n
`
`FIGURE 3
`
`

`

`
`
`US.
`-1
`
`s1ie¢t4ots
`Jul. 10,1990
`Patent
`4,940,835
`GAATTCCCTCAAICTTTACTTTcAAGAATGGctcAAATTAAcAACATGGCTCAAGGGATA
`HeIAlIGInlleA:nAsnHeIAInfilnslyxle
`
`CAAACCCTTAAICCCAATTCCAAITICCATAAACCCCAAGTICCTAAATCTICAAGTTTI
`GInThrL:uAsnProAsnSeIAsnPheHIsL1sProGInV|IPvoLysserseI5etPhe
`
`IN
`
`cT1GTTTT1GGATcTAAAAAACTGAAAAATICAGCAAATTCTAIGTTGGTTTIGAAAAAA
`LeuvalPhealy$orLy|LysLeuLyaAsnSerAlIlsnserIoILeuV||leuLysLys
`
`1M
`
`GATTCAATTTTTAIGCAAAAGVTTTGTTCCTTYAGGAITTCAGCATCAGTGGCTACAGCA
`AspserlloPholI|GluLysPheCyssetPhuArglIeserAlaservnlllnlhraln
`I---amature EPSPS
`-
`
`CAGAAGCCITCTGAGATAGTGTTGCAACCCATTAAAGAGATTICAGGCACTBTTAAATTG
`GInLysPIo5orGlulleVnlLnuGlnProIlIlysclulIoSevGlyIhrV:ILysL|u
`
`CCTGGCTCIAAATCATTATCTAATAGAATTCTCCTTCTTGCTGCCTTATCTGAAGGAACA
`ProfilyserlyssotlonsorA:nArgI|oLcuLeuLuuAlaAlllouserfilnalyfhr
`
`ACTGTGGTTGACAATIIACTAAGTAGTGATGATATICATTACATGCTTGGTGCCTTGAAA
`ThrVa|VulAsphlnleulenSelserlsplqplleqisTyrIpIleuG|yA|aLcuLys
`ACACTTGGACTGCLTGTAGAAGAAGATAGTGCAAACCAACGAGCTGTTGTTGAAGGTTGT
`ThrleuslyleuflisvalfilnGIuAspSerA|aAsnGlnArgAlavalvalelnfilycys
`
`GGTGGGCTTTTCCCTGTIGGTAAAGAGICCAAGGAAGAAAITCAACTGYTCCTTGGAAAT
`GlyGIyLeuPhePvoV:IG|1LysGIu5etLysGluGlulleGlnLeuPh:L:uGlyAsn
`
`GCAGGAAQAGCAATGCGGCCACTAACIGCAGCAGTIACTGTAGCTGGIGGAAATTCAAGG
`AlnGlyThtAIaIe1ArgPrnLeuThrAl|AlaVnIThrValAlaelyfilylsnserkrg
`
`HM
`
`TATGTACTTGATGGAGTTCCTCGAATGAGAGAGAGACCAAITAGTGAITTGGTTGAIGGT
`TyvVaILouAspGIyV:IIrakuglutkrgsluArgProlIeSIrAspLeuValAspGIy
`
`CTTAAACAGCTTBGTG¢AGAGGTTGATTBTT1CcTTGGTAC6IAATGTCCTCCTGTTCGA
`LonLysGlnLcuBIyAI|G|uvaIAsp¢ysPheLeuGlyThuLy:CysPtoProV|lAIg
`
`ATTGTCAGCAAGGGAGGTCTTCCIGGAGGGAAGGTCAAGCTCTCTGGATCCATTAGCAGC
`IlovnlsnvtysclyclylenPraclyelyLy|V:|LysLnnSerGlyselllesevser
`
`CAATACTYGACTGCYCTGCTTATGGCTGCTCCACTGGCTTYAGGAGATGTGGAGATYGAA
`GInTyvLInTitA||LouLouIelAlaAlnProLonAlaLouGlyAspV|IGIulleslu
`
`ATCATTGACAAACTAAITAGTGYACCTTATGTCGAGATGACATYGAAGTTGATGGAGCGA
`llolleksplyslunllnsctVaIProTyrValGIuIo1IhrLeuLysLauIetGIuArg
`
`TTTGGTAITTCYGTGGAGCACAGTAGTAGCTGGGACAGGITCTTTGTCCGAGGAGGTCAG
`FItGlyIlnstrvllalufllssIrsorS|tTrpAspAtgPhoPheVaIArgGlyGIyGIn
`
`AAATACAAGTCTCCTGGAAAAGCTTIYGTCGAAGGTGATGCTTCAAGTGCTAGCTACTTC
`Ly:Tyrly:5erPIo6|ylysAI|PIeVaIGlue!yhxpllaserserllnscrTyrPhe
`
`TTGGCTGGTGCAGCABYCACAGGTGGAACIATCACTGTTGAAGGTTGTGGGACAAACAGT
`LeuAlaGIyAI|AlaVaIThrGIyGIyThr|lIThtV|lGlnalycysfilyThrA:nSer
`
`TTACAGGGGGATGTCAAATTTGCTGAGGTACTTGAAIAAATGGGAGCIGAAGIIACGIGG
`LtuclnelyktpvalLysPIeAl|GluV|ILouG|uLysIalG!yAl|GluV|IThrIrp
`
`Wfl
`
`INN
`
`WW
`
`Ifih
`
`NW
`
`HGURE4a
`
`

`

`
`
`US. Patent
`
`Jul. 10, 1990
`
`Sheet 5 of 8
`
`1141
`
`1EH
`
`1flfl
`
`flfl
`
`1flh
`
`HM
`
`EN
`
`1%“
`
`W5
`
`Ififl
`
`WM
`
`INN
`
`1H
`
`1§H
`
`ACAGAGAACAGTGTCACAGTCAAAGGACCTCCAAGGAGTTCTTCTGGGAGGAAGCATTTG
`ThtGluAsnSarV|I1hrV|lLysGlyPtoProArgS:rSIr5e!GlyArgLysHlsLeu
`
`4,940,835
`1D
`
`cGTGcCATTGATGTGIAcATGAATAAAATGCCTGATGTTGCCATGACACTTGCIGITGTT
`ArgA|alIeAspValksnHelAsnLysHelProAspValAIaflelThrLnuA|Ivalvu
`
`GCACTTTATGCTGATGGTCCCACAGCTATAAGAGATGTTGCTAGCTGGAGAGTCAAGGAA
`AIILenTyrAI|AspGlyProThtA|al|eArgAspValAl|Set1rpArgVaILysGIb
`
`ACTGAGCGCATGATCGCCATATGCACAGAACTTAGGAAGTTAGGAGCAICCGTTGAAGAA
`TnrGIuArgI:1II:AlallacysThrGluLeuAIgLysLenGlyhlathrvnlGIunlu
`
`GGACCAGACTACTGCATAATCAcC¢CAcCGGAGAAAcTAAAtGTGACCGATATTGATACA
`GlyPtoAspTyrcyslIclIuThtProFroG|uLysLauAsnVnlIhrAspIleAsp!hr
`
`TAcGATGATcAcAGGAATGccATGGCTTTTTCTCTTGCTGCTTBTGCAGAIGTTCCCGTC
`TyrtsplspflisArgAsnAI:HeIAI:PheSetleuAlaA|acysAIaAspV:IProVn
`
`ACCATCAAYGACCCTGGCTGCACGCGGAAAACCTTCCCTAACTACTTTGATGTACTTCAG
`ThrlI|AsnAspPvoGIycysTh:ArgLysThrPh:ProAsnTyr?heAspvalLeucln
`
`CAGTACTCCAAGCAITGAACCGCTTCCCTATATTGCAGAATGTAAGTAAGAA1ATGIGAA
`GInTyISutLysHisEnd
`
`GAGTTTAGTICTTGTACAAGACAGGCTACGACTGCCTGGTATCAGAACCICAAIGGGTTB
`
`CATTTCAGTTC‘GIAGGGClTTCCAAGGCTTCGAACTQTTTACTTATTTGCGAGTGATGA
`
`AATGTATTTGTTAGAGTTGAGCTTCTTTTTGTCTTTAAGGAATGYACACTAATAGAGTIA
`
`AGAATIACTIGTATGGGCCAGTGTAAGGAGTACTATTACTCTTTGCTTATTTTATTGATT
`
`GAGTTTYGTCAAGGATCTGGCTTTG7CAAGAATTACTGGT+AATTTTATFGACAATCTCA
`TGTGICTAAATGAAATTGTTTGITAAAAAAAAAAAAAAAAIAAKAAAAAAAGGAAITE
`
`13W
`
`flfl
`
`1&0
`
`an
`
`E3
`
`1N»
`
`nw
`
`1D
`
`WE
`
`1ND
`
`IN?
`
`Esufl
`
`Enfl
`
`I-nfll
`
`Illflll
`
`Ital
`
`51:03
`
`0
`
`IN
`
`IND
`
`HGURE4b
`
`

`

`
`
`U.S. Patent
`
`JuL 10, 1990
`
`Sheet 6 of 8
`
`4,940,835
`
`RIGHT BORDER
`
`FIGURE 5
`
`

`

`US. Patent
`
`J
`
`8.m.
`
`4,940,835
`
`:1...=......E8:0Eng8=3:.:3:__._nlM83Z02mama202..53205..
`
`..\..IS.........r...
`
`
`.......8:8-.8».Wfl.n::...n..:.3.
`
`
`
`
`7Aat8:M:3.Easy=.u...x.......I.!.=!..::25::3:=:-an.EEu:5.
`
`
`9.=!..:
`3.53.8u.
`
`......................
`ao\-I
`
`........./..
`,,m_maou_aE<
`
`m_..==on_
`
`0mmaum
`
`._.._3:u3u_n:n.=:=U.
`
`coxwl_________
`
`
`
`
`

`

`US. Patent
`
`Jul. 10, 1990
`
`Sheet 8 of 8
`
`4,940,835
`
`pMON9721 (12Kb)
`
`RIGHT BORDER
`
`FIGURE 7
`
`

`

`
`
`1
`
`GLYPHOSATE-RESISTANT PLANTS
`
`4,940,835
`
`2
`pet-iments have demonstrated that such a method would
`be efiicacious in difieraitiated plants.
`Aiher the filing date of U.S. Pat. No. 4,53 5,060, meth-
`ods and vectors were described which could-be used to
`insert foreign genes into plant cells (see, e.g.. Fraley
`1983, Herrera-Estrella I983. Bevan I983, and PCT ap-
`plications W0 No. 84/02919 and 02920). in PCT appli-
`cation WO No. 84/02913. methods were also described
`for creating
`having bacterial EPSPS
`coding sequences controlled by regulatory seqnencs
`derived from genes which are active in plant cells.
`Using these vectors and methodology, bacterial genes
`such as the mutant Salmonella EPSPS gene mentioned
`abovecanhemanipulatedandexpressedhiplanteells.
`Theobject of this invention is to provide a method of
`genetically transforming plant cells which causes the
`cellsandplantsregeneratedtherefromtobwomeresis
`tant to glyphosate and the herbicidal salts thereof.
`SUMMARY OF THE INVENTION
`
`This application is a Continuation-in-Part of applica-
`tion, Ser. No. 792.390 filed Oct. 29, 1985. now aban-
`doned. which. in turn, is a Continuation-in-Part of appli-
`cation. Ser. No. 763,482, filed Aug. 7, 1985, now aban-
`cloned.
`
`BACKGROUND OF THE INVENTION
`
`The present invention relates to the fields of genetic
`engineering, biochemistry, and plant biology.
`N-phosphonomethylglycine has the following struc-
`ture:
`
`OH
`
`O
`H
`ll/
`I
`I
`HO-C-CBz—N-CH;-P
`
`0I
`
`OH
`
`Thismo1eculeisnnneid,whichcandissociateinaque-
`ous solution to form phytotoxicant anions. Several ani-
`onicformsareknown.Aausedherein,thename“gly-
`phosate"re.ferstotheacidanditsanions.Ami::ture
`containing flyphosate as the active ingredient, formu-
`latedasitsisopropylaminesalt,issoldasaherbicideby
`Monsanto Company under the trademark ROUN-
`DUI’®. Numerous other salts also have herbicidal
`properties, as exemplified by U.S. Pat. No. 3,799,758
`(Franz I974) and various other patents. Compositions
`comprising N-phosphonomethylglycine and salt-form-
`ing cations which increase the solubility of the N-phos-
`phonomethylglydne in water are preferred.
`Ihosesldnedintheanrecogziizethatthescientific
`literature contains numerous papers suggesting several
`modesofactionforinhihitianofplantgrowthbygly-
`phosate.
`proposed mode suggsts that glyphosate
`inhibits an enzyme called 5-cnolpyruvylshilu'mate-3-
`phosphate synthase (I-ZPSPS); see, e.g., Amrhein 1980,
`Steinrucken 1980. Mousdale 1984, and Rubin 1982
`(note: a complete fat of references is contained below,
`after the Examples). The EPSPS enzyme reportedly
`catalyzes the conversion of shildmate-3-phosphntc into
`S-enolpyruvyl-shikimate-3-phosphate. an intermediate
`in the biochemical pathway for creating three essential
`aromatic amino acids (tyrom phenylalanine, and
`tryptophan); see, e.g., Mousdale i984. Rogers 1983
`reports that overproduction of EPSPS in E colt‘ con-
`tributes to glyphosste ruistance in those cells.
`Atleastoueresearcherhasattemptedtocreate gly-
`phosate-resistanthactuialceflshymnnipuladngabacte
`rialgene whieh'encodesanEPSPS enzyme. As de-
`scribed in Us. Pat. No. 4,535,060 (Comai; assigned to
`Calgene. Inc.;filingdateJan. 5, l933)andin Comai
`1983, a culture of Salmonella bacteria was contacted
`with amutagen(ethylmethanesuifonate). Thebacteria
`wu’escreenedforg‘lyphosateresistauceandare1a-
`tivelyresistantculturewasselected.Thiscultnrewas
`analyzed. and determined to have a mutant form of
`EPSPSwithasubstitutedaminoacid.ureportedin
`Stalker 1985. U.S. Pat. No. 4,535,060 suggested that the
`mutnntEPSPSgenecouJdbeinsertedintoplnntcellsto
`create glyphosnte-resistant (Glyk) plant cells. In addi-
`tion, ithasbeenreportedthntglyphosatetolernmplant
`cellscanbeselectedwhiohoverproduceEPSPSinthe
`preaenceoflowlevelsofglyphosate(l*Ia.fzigereta1.
`1984 and Smart et al, 1985). However, none of the ex-
`
`30
`
`35
`
`45
`
`50
`
`60
`
`65
`
`This invention involve a cloning or expression vec-
`tor comprising a gene which encodes 5-enolpyruvyl-
`shilrimate-3-phosphate synthase (EPSPS) polypeptide
`which, whenexpressedinaplantcellcontainsaehloro-
`plast transit peptide which allows the polypeptide, or an
`enzymaticallyactiveporfionthereofitobetransported
`fmmthecytoplasm oftheplantcellintoachlcroplastin
`the plant cell, and confers a substantial degree of gly-
`phosnte resistance upon the plant c