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* Confidential Treatment Requested
EXHIBIT 10.15
RESEARCH AGREEMENT
THIS RESEARCH AGREEMENT ("Agreement") is made this 31st day of August,
1998, by and between Third Wave Technologies, Inc. ("Institution"); and
Xxxxxx-Xxxxxxx Company, a Delaware corporation, with its principal place of
business at 000 Xxxxx Xxxx, Xxxxxx Xxxxxx, Xxx Xxxxxx, 00000 ("Company").
RECITALS
1. Institution has developed substantial expertise in the area of assay
development for gene expression profiling and polymorphism and mutation analysis
(the "Field").
2. Company desires to obtain the services of Institution in directing certain
research in the Field relating to the development of certain assays. Such
research is hereinafter referred to as the "Development Program". Institution is
willing to direct the Development Program, all on the terms and conditions set
forth herein and in accordance with the protocol attached hereto as Exhibit A
and incorporated herein by reference (the "Protocol").
NOW, THEREFORE, in consideration of the covenants and premises herein
contained, the parties agree as follows:
Section 1. The Development Program. Promptly following receipt of the
amount set forth in Section 2 below, Institution shall use diligent efforts to
develop the Invader(R) assay for the five (5) analytes as described in the
Protocol, subject to Company providing Institution the materials described in
Exhibit B (collectively, the "Company Materials"). Upon completion of the
Development Program with respect to each analyte, Institution agrees to provide
enzyme buffer and Invader probes reasonably sufficient for Company to perform
five hundred (500) assays for such analyte (collectively, the "Institution
Materials"). Accordingly, Institution shall, after consultation with Company,
direct and perform the Development Program in a professional and diligent
manner, all in accordance with the Protocol and the terms of this Agreement.
Section 2. Payment. In consideration for the performance of the Development
Program by Institution, Company shall pay Institution a non-refundable amount of
[****] (the "Research Amount") within five (5) days of the date of Company's
execution of this Agreement.
Section 3. Reporting. Company designates Xxxxxxx Xxxx or his or her
successor to function as its research and development contact (the "Contact").
Institution shall inform the Contact of the progress of the Development Program
in the following manner:
a. By informal verbal reports, from time to time;
b. By response to all of Company's reasonable inquiries regarding the status of
the Development Program;
c. By arrangements to periodically meet and discuss the progress of the
Development Program with the Contact at Institution's facilities; and
d. By submission of a detailed written report thirty (30) days following the
conclusion of the Development Program (the "Final Report").
Section 4. Ownership of Data: Publication. Subject to Company's rights in
the Company's Materials, all right, title and interest in and to all information
disclosed by Institution hereunder and to all
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Institution Materials transferred hereunder shall remain vested in Institution.
Except as otherwise provided in this Section 4 below, Company shall own all
right, title and interest in and to Inventions (collectively, "Company
Inventions") and Institution hereby assigns all such right, title and interest
to Company. Furthermore, Institution hereby agrees to only use its personnel in
connection with the Development Program. Notwithstanding the foregoing, Company
hereby agrees that Institution shall own all right, title and interest in and to
Inventions directed to subject matter comprising improvements to the Invader
assay technology, including without limitation methods of performing Invader
assays (collectively, "Improvements") and Company hereby assigns all such right,
title and interest to Institution. In addition, to the extent that it has the
right to do so Company hereby grants to Institution an exclusive, worldwide,
royalty-free, fully paid-up, irrevocable right and license, under any and all
Company Inventions, to make, have made, use, import, offer for sale and sell
products and components, solely for diagnostic applications including for tests
or assays used to guide, prescribe or direct therapeutic regimens, treatments or
drugs. For purposes of this Agreement, "Invention" shall mean any and all
discoveries, inventions and other subject matter (whether patentable or not)
made in the course of performing the Evaluation or otherwise in connection with
using the Institution Materials as permitted hereunder and all intellectual
property rights therein. Nothing in this Agreement is to be construed as
granting a license to Company to utilize any information received from
Institution or Institution Materials except as expressly provided herein, under
any patent or other intellectual property rights owned by Institution, unless a
separate agreement for such rights is executed by Company and Institution.
Company and Institution further agree to perform such acts and provide such
documents as are reasonably requested by the other party to effect the foregoing
assignments.
Section 5. Confidentiality.
(a) Confidential Information. The parties may from time to time
disclose to each other Confidential Information. "Confidential Information"
shall mean and information disclosed by one party to the other party hereto
which if disclosed in tangible form is marked "confidential" or with other
similar designation to indicate it's confidential or proprietary nature or if
disclosed orally is indicated orally to be confidential or proprietary by the
party disclosing such information at the time of such disclosure and is
confirmed in writing as confidential or proprietary by the disclosing party
within a reasonable period after such disclosure; provided, however, all
Improvements shall be deemed to be Confidential Information of the Institution
and all Company Inventions (except to the extent that such Company Inventions
related to diagnostic applications) shall be deemed to be Confidential
Information of the Company. Notwithstanding the foregoing or anything herein to
the contrary, Confidential Information shall not include any information that,
in each case as demonstrated by written documentation: (i) was already known to
the receiving party or its affiliates, other than under an obligation of
confidentiality, at the time of disclosure; (ii) was generally available to the
public or otherwise part of the public domain at the time of its disclosure to
the receiving party; (iii) became generally available to the public or otherwise
part of the public domain after its disclosure and other than through any act or
omission of the receiving party in breach of this Agreement; or (iv) was
subsequently lawfully disclosed to the receiving party or its affiliates by a
third party which is not in violation of any contractual or legal obligation to
the disclosing party with respect to such Confidential Information.
(b) Confidentiality. Each party agrees to hold and maintain in strict
confidence all Confidential Information of the other party. Without limiting the
foregoing, neither party shall use or disclose the Confidential Information of
the other party, except as otherwise permitted by this Agreement or as may be
necessary or useful to exercise its rights or perform its obligations under this
Agreement. Nothing contained in this Section 5 shall prevent either party from
disclosing any Confidential Information of the other party (1) to accountants,
lawyers or other professional advisors to the extent reasonably necessary to
accomplish the purposes of this Agreement or to prospective lenders, investment
bankers or other financial institutions in connection with a merger, acquisition
or securities offering, subject in each case to the recipient entering into an
agreement to protect such Confidential Information from disclosure; or (ii) to
the extent is required by law or regulation to be disclosed; provided, however,
that the party subject to such disclosure requirement has provided written
notice to the other party
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promptly upon receiving notice of such requirement in order to enable the other
party to seek a protective order or otherwise prevent disclosure of such
Confidential Information. Upon any termination of this Agreement, Company and
Institution shall promptly return to the other party all Confidential
Information received from the other party (except one copy of which may be
retained for purposes of monitoring compliance with the provisions of this
Section 5 and for archival purposes).
The foregoing provisions of this Section 5 shall survive expiration or
termination of this Agreement for a period of seven (7) years after such
expiration or termination.
Section 6. Compliance with Applicable Laws. Institution agrees to conduct
the Development Program and maintain records and data during and after the term
of this Agreement in compliance with all applicable legal and regulatory
requirements, including without limitation any applicable requirements of the
United States Food and Drug Administration and the United States Federal Drug
Enforcement Administration.
Section 7. Materials
(a) Company agrees that all Institution Materials obtained from
Institution pursuant to this Agreement shall be used solely for the purpose of
evaluating whether or not Company desires to enter into a licensing and other
business arrangements with Institution, all on terms and conditions as mutually
determined by Company and Institution and not for any commercial purposes (the
"Evaluation"). Institution agrees that all Company Materials obtained from
Company pursuant to this Agreement shall be used solely for the purpose of
evaluating whether or not Institution desires to enter into a licensing and
other business arrangements with Company, all on terms and conditions as
mutually determined by Company and Institution and not for any commercial
purposes other than the Development Program and commercial diagnostic
applications. Company agrees at all times to use the Institution Materials, and
Institution agrees at all times to use the Company Materials, in compliance with
all state, federal and other applicable laws, rules and regulation pertaining to
use thereof. The Institution Materials and the Company Materials shall include
the original materials transferred to Company or Institution, respectively, as
well as any derivatives or improvements developed by the receiving party
therefrom. INSTITUTION SUPPLIES THE INSTITUTION MATERIALS AND COMPANY SUPPLIES
THE COMPANY MATERIALS WITHOUT ANY WARRANTY, REPRESENTATION OR UNDERTAKING
WHATSOEVER, EXPRESSED OR IMPLIED, INCLUDING, BUT NOT LIMITED TO, ANY WARRANTY
RESPECTING THE EFFICIENCY, PERFORMANCE, WORKMANSHIP, CONDITION, MERCHANTABILITY,
FITNESS FOR PARTICULAR PURPOSE OR NONINFRINGEMENT.
(b) During the term of this Agreement and for five (5) years
thereafter Institution agrees to make available to Company for its own internal
research and development purposes enzyme buffer and Invader probes developed
hereunder pursuant Institution's standard terms and conditions therefor,
including standard pricing and minimum order requirement of at least quantities
to perform one thousand (1,000) assays for any particular analyte.
Section 8. Institution's Representations and Warranties. Institution hereby
represents, warrants and covenants to Company the following:
a. It has the power and authority to undertake the contractual commitments
set forth in this Agreement.
b. It is free to enter into this Agreement and carry out its obligations
hereunder without violating any obligation owed to a third party, including,
without limitation, any governmental or quasi-governmental agency, group or
department, or any other private or public institution, person or company. No
such third party currently has, or, except as expressly authorized herein, will
have, any option, license
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or other right of any kind with respect to any Study Drug or any data,
information, inventions or discoveries obtained or developed under this
Agreement.
Section 9. Monitoring of Development Program. During the term of this
Agreement, Institution agrees to permit representative of Company to examine at
any reasonably time during normal business hours (i) the facilities where the
Development Program is being conducted, (ii) raw research data and (iii) any
other relevant information (and to make copies) necessary for Company to
confirm that the Development Program is being conducted in conformance with the
Protocol and in compliance with applicable laws and regulations, including
those of the United States Food and Drug Administration and the United States
Federal Drug Enforcement Administration.
Section 10. Term. The term of this Agreement shall commence on the date
that it is made and continue for up to 90 days.
Section 11. Termination. The Development Program may be terminated by
Company at any time in the exercise of its sole discretion upon fifteen (15)
days prior written notice to Institution. Upon receipt or giving of notice, as
the case may be, Institution agrees promptly to terminate conduct of the
Development Program.
Section 12. Publicity. Neither party shall use the name of the other
party or its divisions, affiliates, personnel or products, as applicable, for
promotional purposes without the prior written consent of the party whose name
is proposed to be used, which consent shall not be unreasonably withheld;
provided, however, that either party may (i) disclose the terms of this
Agreement to prospective lenders, investment bankers and other financial
institutions of its choice solely for the purposes of financing the business
operations of such party, either upon the written consent of the other party or
if the disclosing party obtains a signed confidentiality agreement with such
entity or financial institution with respect to such information, and (ii) make
disclosures to the extent required to comply with applicable securities law and
in the case of (ii), if possible, the non-disclosing party shall have consented
to such disclosure, which consent shall not be unreasonably withheld.
Section 13. Independent Contractor. Institution is acting in the
capacity of independent contractor hereunder and not as employee or agent of
Company.
Section 14. Controlling Law. This Agreement shall be governed by and
construed in accordance with the law of the State of Michigan (other than
provisions relating to conflicts of laws).
Section 15. Agreement Modifications. This Agreement may not be altered,
amended or modified except by written document signed by all parties.
Section 16. Inconsistencies. The terms and conditions of this Agreement
shall govern in the event of conflict between it and the Protocol.
Section 17. Force Majeure. Any party's delay or failure in performing
its obligations under this Agreement shall be excused to the extent caused by
the occurrence of events beyond that party's reasonable control, including
without limitation, war, floods, earthquakes, other acts of God, industrial
disputes, civil disobedience, strikes, fire, mobilization, changes in
governmental regulation or interpretation, requisition, embargo, restriction
and shortage of transport facilities, fuel, energy or supplies. Each party
claiming the benefit of such an excuse shall notify the other party in writing
of any such delay or failure in performance, and shall resume performance as
soon as is reasonably practicable.
Section 18. Notice. All notices given hereunder shall be in writing and
shall be delivered by hand or mailed by certified or registered mail, return
receipt requested, postage pre-paid, addressed to the parties as follows:
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To Company:
Xxxxx-Xxxxx Pharmaceutical Research
0000 Xxxxxxxx Xxxx
Xxx Xxxxx, Xxxxxxxx 00000
Attn: Chairman
with a copy to:
Xxxxx-Xxxxx Pharmaceutical Research
0000 Xxxxxxxx Xxxx
Xxx Xxxxx, Xxxxxxxx 00000
Attn: Assistant General Counsel
To: Institution:
Third Wave Technologies, Inc.
000 Xxxxx Xxxx Xxxx
Xxxxxxx, Xxxxxxxxx 00000-0000
Attn: President
Notice shall be deemed given when received and the parties may change the
respective addresses where notice is to be given upon prior notice hereunder.
IN WITNESS WHEREOF, the parties hereto have caused this Agreement to be
executed by their duly authorized representatives as of the date first above
written.
XXXXXX-XXXXXXX COMPANY
By: /s/ XXXXXXX XXXXXXXX, Ph.D.
-----------------------------
Name: Xxxxxxx Xxxxxxxx, Ph.D.
----------------------------
Title: Senior Vice President, Worldwide
Preclinical Research, Development and Technologies
Xxxxx-Xxxxx Pharmaceutical Research
THIRD WAVE TECHNOLOGIES, INC.
By: /s/ XXXXXXX TREBLE
-----------------------------
Name: Xxxxxxx Treble
Title: Chief Operating Officer
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EXHIBIT A
PROTOCOL
The Institution will develop five (5) discrete assays as follows:
Three (3) Gene Expression Assays as defined in Exhibit B for each of three
(3) unique mRNA targets as specified by the Company. The Gene Expression
Assays will utilize the Institution's Primary Invader Technology and
detection in the polymerase/streptavidin-coated microtiter plate format.
Two (2) Polymorphism and Mutation Assays as defined in Exhibit B for each
of two (2) unique single nucleotide polymorphisms as specified by the
Company. The Polymorphism and Mutation Assays will utilize the
Institution's Secondary Invader Technology and detection in the microtiter
plate format using Fluorescence Resonance Energy Transfer (FRET).
A. The assay Development Program for each Gene Expression Assay will encompass
the following:
1. Prior to initiation of the Development Program, the Company will provide
the Institution with the appropriate genetic sequence specifying the
mRNA target of interest in Exhibit B.
2. The Institution will design two target-specifying oligonucleotides (an
"Invader" oligo and a "Signal" probe) for use in the Primary Invader
reaction. The oligonucleotides will be designed to hybridize to the
target such that 3'-end of the Invader oligo will overlap the 5'-most
region of hybridization of the Signal probe. The Institution's
proprietary Cleavase enzymes are designed to recognize the structure
created by this overlapping region and cleave the 5' end of the Signal
probe for subsequent detection.
3. The melting temperature (T(m)) of the target-specific Signal probe will
be estimated. In order to optimize signal generation, a temperature near
the T(m) of the Signal probe will be utilized for the Primary Invader
reaction such that the reaction cycle (hybridization of the Signal
probe, cleavage of the 5' end of the Signal probe, release of the
remaining 3' portion of the Signal probe) will occur rapidly under
isothermal conditions. This rapid isothermal cycling allows each copy of
target mRNA to serve as the substrate for multiple signal generation
cleavage events during reaction incubation. The accumulation of signal
(5' fragments of the Signal probe) is directly proportional to the
number of target mRNA transcripts present in the test sample.
4. The Institution will design a biotin labeled oligonucleotide for capture
of accumulated Signal probe fragments on a streptavidin-coated
microtiter plate. The complex formed by capture of the Signal probe
fragments on the plate forms a primer-template substrate for DNA
polymerase which is used to extend the Signal probe fragment with
fluorescein-dUTP (FdUTP).
5. The Institution will select an appropriate anti-fluorescein alkaline
phosphatase-conjugated antibody and chemifluorescent substrate for
detection of the FdUTP labeled fragments.
6. The Institution will develop appropriate standards for use in the assay
and will demonstrate assay proof-of-principle using this material. From
time to time during the assay development program, the Company will
provide the Institution with appropriate cellular material known to have
the mRNA target of interest expressed at various levels. The Institution
will use this material for further assay refinement and optimization.
7. The Institution estimates that a minimum of 10 attomoles of the mRNA
target will be required in each test sample for detection in the assay.
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B. The assay Development Program for each Polymorphism and Mutation Assay will
encompass the following when the starting sample material is a PCR product or
Genomic DNA:
1. At the initiation of the Development Program, the Company will provide
the Institution with the appropriate PCR product and Genomic DNA
containing the target of interest.
2. Primary Invader Reaction:
- [****] The oligonucleotides will be designed to hybridize to
specific targets (wild-type or mutant respectively) such that 3'-end
of the Invader oligo will overlap the 5'-most region of
hybridization of either the wild-type or mutant probe. The
Institution's proprietary Cleavase enzymes are designed to recognize
the structure created by this overlapping region and cleave the 5'
end of the wild-type and mutant probes for use in the Secondary
Invader reaction.
- The melting temperature (T(m)) of both the wild-type and mutant
probes will be estimated. In order to optimize signal generation, a
temperature near the T(m) of these probes will be utilized for the
Primary Invader reaction such that the reaction cycle (hybridization
of probes, cleavage of the 5' end of the probes, release of the
remaining 3' portion of the probes) will occur rapidly under
isothermal conditions. This rapid isothemal cycling allows each copy
of PCR product or Genomic DNA target to serve as the substrate for
multiple probe cleavage events during reaction incubation. The
accumulation of wild-type arms (5' fragments of the wild-type probe)
is directly proportional to the number of wild-type target molecules
present in the test sample. The accumulation of mutant arms (5'
fragments of the mutant probe) is directly proportional to the
number of mutant target molecules present in the test sample.
3. Secondary Invader Reaction:
- [****] of the wild-type and mutant probes from the Primary Invader
reaction are designed to act as Invader oligonucleotides in the
Secondary Invader reaction. The oligonucleotides will be designed
such that the 3'-end of the Invader oligos (from the Primary Invader
reaction) will overlap the 5'-most region of hybridization of the
Signal probe to the secondary target oligo. The Institution's
proprietary Cleavase enzymes are designed to recognize the structure
created by this overlapping region and cleave the 5' end of the
Signal probe for subsequent detection. [****]
- The melting temperature (T(m)) of the Signal probe will be
estimated. In order to optimize signal generation, a temperature
near the T(m) of the Signal probe will be utilized for the Secondary
Invader reaction such that the reaction cycle (hybridization of
Signal probe, cleavage of the 5' end of the Signal probe, release of
the remaining 3' portion of the Signal probe) will occur rapidly
under isothermal conditions. This rapid isothemal cycling allows
each copy of secondary target to serve as the substrate for multiple
signal generation cleavage events during reaction incubation. The
accumulation of signal (5' fragments of the Signal probe) is
directly proportional to the number of Secondary Invader oligos
(from the Primary Invader reaction) present in the secondary
reaction.
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4. The Institution will determine appropriate assay cut-off values for
assessment of genotype (wild-type homozygous, mutant homozygous,
wild-type/mutant heterozygous) based on fluorescence signal.
5. The Institution estimates that a minimum of 20ng of PCR product or 100ng of
Genomic DNA will be required in each test sample for detection in the assay.
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EXHIBIT B
DEVELOPMENT OF THREE (3) GENE EXPRESSION ASSAYS
Company has identified three unique mRNA targets for which Institution will
develop Gene Expression assays as specified below.
1) [****] The sequences are very similar. Invaders should be directed to regions
of identity to these sequences.)
LOCUS [****]
DEFINITION [****]
ACCESSION [****]
NID [****]
KEYWORDS [****]
SOURCE [****]
ORGANISM [****]
Eukaryotae; mitochondrial eukaryotes; Metazoa; Chordata;
Vertebrata; Eutheria; Rodentia; Sciurognathi; Myomorpha; Muridae;
Murinae; Rattus
REFERENCE 1 (bases 1 to 813)
AUTHORS Xxxxxxxx,X.X., Xxxxx,X.X., Xxxxxxx,X.X., Xxxxx,X.X.,
Xxxxxxxx,X.X., Xxxx,X.X. and Xxxxxxxxx,P.
TITLE "Cloning and sequence of several alpha-2u-globulin cDNAs"
JOURNAL Proc Natl. Acad. Sci. U.S.A. 78, 3478-3482 (1981)
MEDLINE 81273082
COMMENT Amino acid sequence encoded by bases 46 thru 135 was
independently determined.
FEATURES Location/Qualifiers
source 1..813
/organism="Rattus norvegicus"
/db_xref="taxon:10116"
mRNA <1..813
/note-"a-2u mRNA"
CDS <1..534
/note="alpha-2u globulin precursor()"
/codon_start=1
/db_xref="PID:g204261"
/translation="LLLLCLGLTLVCGHAEEASSTSGNLDVAKLNGDWFSIVVASNKREKIEENGSMRVFM
QHIDVLENSLGFKFRIKENGECRELYLVAYKTPEDGEYFVEYDGGNTFTILKTDYDRYVMFHLINF
KNGETFQLMVLYGRTKDLSSDIKEKFAKLCEAHGITRDNIIDLTKTDRCLQARG"
sig_peptide 4..45
/note="alpha-2u globulin signal peptide"
mat_peptide 46..531
/note="alpha-2u globulin"
BASE COUNT 233 a 180 c 185 g 215 t
ORIGIN 38 bp upstream of BalI site.
1 ctgctgctgc tgtgtctggg cctgacactg gtcgtggcc atgcagaaga agctagttcc
61 acaagcggga accctcgatgt ggctaagctc aatggggatt ggttttctat tgtcgtggcc
121 tctaacaaaa gagaaaagat agaagagaat ggcagcatga gagtttttat gcagcacatc
181 gatgtcttgg agaattcctt aggcttcaag ttccgtatta aggaaaatgg agagtgcagg
10
241 gaactatatt tggttgccta caaaacgcca gaggatggcg aatattttgt tgagtatgac
301 ggagggaata catttactat acttaagaca gactatgaca gatatgtcat gtttcatctc
361 attaatttca agaacgggga aaccttccag ctgatggtgc tctacggcag aacaaaggat
421 ctgagttcag acatcaagga aaagtttgca aaactatgtg aggcgcatgg aatcactagg
481 gacaatatca ttgatctaac caagactgat cgctgtctcc aggcccgagg atgaagaaag
541 gcctgagcct ccagtgctga gtggagaact tctcaccagg actctagcat caccatttcc
601 tgtccatgga gcatcctgag acaaattctg cgatctgatt tccatcctct gtcacagaaa
661 agtgcaatcc tggtctctcc agcatcttcc ctagttaccc aggacaacac atcgagaatt
721 aaaagctttc ttaaatttct cttggcccca cccatgatca ttccgcacaa atatcttgct
781 cttgcagttc aataaatgat taccttgca ctt
2) [****]
LOCUS [****]
DEFINITION [****]
[****]
ACCESSION [****]
NID [****]
KEYWORDS
SOURCE [****]
ORGANISM [****]
Eukaryotae; mitochondrial eukaryotes; Metazoa; Chordata;
Vertebrata; Mammalia; Eutheria; Rodentia; Sciurognathi; Muridae;
Murinae; Rattus.
REFERENCE 1 (bases 1 to 1182)
AUTHORS Pataon, V.G., Xxxxxxxxxxx, X.X. and Xxxxxxx, X.X.
TITLE "Cloning and subcellular localization of hamster and rat
isopentenyl diphosphate dimethylallyl disphosphate isomerase.
A PTS1 motif targets the enzyme to peroxisomes
JOURNAL J. Biol. Chem. 272, 18945-18950 (1997)
REFERENCE 2 (bases 1 to 1182)
AUTHORS Xxxxx, X.X., Xxxxxxxxxxx, X.X. and Xxxxxxx, X.X.
TITLE Direct Submission
JOURNAL Submitted (12-MAY-1997) Biology, San Diego State University,
0000 Xxxxxxxxx Xx., Xxx Xxxxx, XX 00000, XXX
FEATURES Location/Qualifiers
source 1..1182
/organism="Rattus norvegicus"
/strain="Xxxxxxx-Xxxxxx"
/db_wref="taxon:10116"
CDS 386..1069
/EC_number="5.3.3.2"
/note="IPP isomerase; localized in peroxisomes"
/codon_start=1
/product="isopentenyl diphosphate:dimethylallyl
diphosphate isomerase"
/db_xref="PID:g 2253701"
/translation="MPEINASNLDEKQVQLLAEMCILIDENDNKIGADTKKNCHLNENIDKGLIHRAFSVFL
FNTENKLLLQQRSDAKITFPGCFTNSCCSHPLNNPGELEENDAMGVKRAAQKRLKAELGIPLEEVD
LNEMNYLTRIYYKAQSDGIWGEHEIDYILFLRKNVTLNPDPNEIKSYCYVSKEELKEILKKEARGEI
KFTPWFKILADAFLFKWWDNLNHLSPFVDHEKIHRM"
BASE COUNT 341 a 237 c 295 g 309 t
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ORIGIN
0 xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx gtgcgggcag tctcagctgt ccactgggag
61 tcgccaacac catctcttgg gctccagggt cgcaatcact accggggatc tccagcaagt
121 cttccggacc cacgcgtaga ggaggtggtg ccaccgcgcg caaacctggc gcggtcgtga
000 xxxxxxxxxx xxxxxxxxxx ggcaattggc tggaggcatg tggcgcggac gggcactggc
241 aggagtgatt ggatcagctc ttaaggggcg gggtctggag gcggagcatg ctgccgagag
301 cgttgaagta cggcgctccg cacagcttct cgacactcct gtctggaatc gctgtgttct
361 aggtcagatc agacattctg tcacaatgcc tgaaatcaat gccagcaatc ttgatgaaaa
421 acaggttcag cttctagcag agatgtgtat tcttattgat gaaaatgaca ataaaattgg
481 ggctgacacc aagaaaaatt gtcacctgaa tgaaaacatt gacaaaggac taatacatcg
541 agctttcagt gtcttcttgt ttaatactga aaacaagctc ctgttacagc agagatcgga
601 tgctaaaatt acctttccag gttgtttcac caatagttgc tgtagtcatc cattaaataa
661 ccccggggag ctggaagaaa atgatgccat gggtgtcaaa cgagcagcac agaagcgctt
721 aaaggcggag ctggggatac ccttggaaga ggttgatcta aatgaaatga attatctaac
781 aagaatttac tacaaggccc aatctgatgg tatctggggt gaacatgaaa ttgattacat
841 tttgtttctg aggaagaatg taaccctgaa tccggatccc aacgagatta aaagctattg
901 ctatgtatca aaggaagaac tcaaagaaat tttgaagaag gaagccaggg gcgaaattaa
961 gtttactcca tggtttaaga ttattgcaga cgcttttctc tttaaatggt gggataactt
1021 aaaccatttg agtccttttg ttgaccatga gaaaatacat agaatgtgaa tgtgtaaatg
1081 atttgagatt tttttctacc tactaagaat ggtttgattt tgttttaaat tgggctccct
1141 tgtatgacac atagatattt tacaaccaga atttgttttg ag
3) [****]
LOCUS [****]
DEFINITION [****]
ACCESSION [****]
NID [****]
KEYWORDS [****]
SOURCE [****]
ORGANISM [****]
Eukaryotae; [****]
Vertebrata; [****]
Murinae; Rattus.
REFERENCE 1 (base 1 to 3275)
AUTHORS Xxxxxxx,X.X.
TITLE Direct Submission
JOURNAL Submitted (17-APR-1990) Xxxxxxx X.X., University of Barcelona,
Unit of Biochemistry, School of Pharmacy, Placa Xxxx XXX, x/x.
00000, Xxxxxxxxx, Xxxxx
REMARK (b 1-38,40,41-62,64-3275)
REFERENCE 2 (bases 1 to 3275)
AUTHORS Ayte,J., Xxx-Xxxxx,G. and Xxxxxxx,X.X.
TITLE "Nucleotide sequence of a rat liver cDNA encoding the cytosolic
3-hydroxy-3-methylglutaryl coenzyme A synthase"
JOURNAL Nucleic Acids Res. 18 (12), 3642 (1990)
MEDLINE 90301491
REMARK (b 1-38,40,41-62,64-3275)
REFERENCE 3 (bases 1 to 3275)
AUTHORS Haegardt,F.G.
TITLE Direct Submission
JOURNAL Submitted (30-JUL-1990) to the EMBL/GenBank/DDBJ database
12
FEATURES Location/Qualifiers
source 1.3275
/organism="Rattus norvegicus"
/strain="Xxxxxxx Xxxxxx"
/db_xref="taxon:10116
/tissue_type="liver"
/clone="lambda-cCS1"
precursor_RNA 1..3275
/note="primary transcript"
CDS 92..1654
/note="cytosolic 3-hydroxy 3-methylglutaryl coenzyme A
synthase (AA 1-520)"
/codon_start=1
/db_xref="PID:g55947"
/db_xref="SWISS-PROT:P17425"
/translation="MPGSLPLNAEACWPKDVGIVALEIYFPSQYVDQAELEKYDGVDAGKYTIGLGQARM
GFCTDREDINSLCLTVVQKLMERNSLSYDCIGRLEVGTETIIDKSKSVKSNLMQLFEESGNTDIEGID
TTNACYGGTAAVFNAVNWIESSSWDGRYALVVAGDIAIYASGNARPTGGVGAVALLIGPNAPVIF
DRGLRGTHMQHAYDFYKPDPDMLSEYPVVDGKLSIQCYLSALDRCYSVYRKKIRAQWQKEGKDKDF
TLNDFGFMIFHSPYCKLVQKSLARMFLNDFLNDQNRDKNSIYSGLEAFGDVKLEDTYFDRDVEKA
FMKASAELFNQKTKASLLVSNQNGNMYTSSVYGSLASVLAQYSPQQLAGKRIGVFSYGSGLAATL
YSLKVTQDATPGSALDKITASLCDLKSRLDSRTCVAPDVFAENMKLREDTHHLANYIPQCSIDSLFE
GTWYLVRVDEKHRRTYARRPSTNDHSLDEGVGLVHSNTATEHIPSPAKKVPRLPATSGEPESAVIS
NGEH"
BASE COUNT 934a 634c 789 g 918 t
ORIGIN
1 cccagggtcc gatcgcgttt ggtgcctgaa ggaggaaccg gtgacagacc tggagactac
00 xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx tcacttcctt tgaatgcaga
121 ggcttgctgg ccaaaagatg tgggaatcgt tgcccttgaa atctactttc cttctcagta
181 tgttgatcaa gctgagttgg aaaaatacga tggtgtagat gctggaaagt ataccattgg
241 cctgggccag gccaggatgg gcttctgcac ggatcgcgaa gacatcaact ctctctgcct
000 xxxxxxxxxx xxxxxxxxxx tggagagaaa tagcctttcc tatgactgca ttggcggct
361 ggaagtcggg acagagacaa tcatcgacaa atcaaaatcc gtgaagtcta atttgatgca
421 gctgtttgag gagtctggga atacagatat agaaggaata gatacaacta atgcatgcta
481 tgggggcaca gccgcagtct tcaatgctgt gaactggatc gaatccagct cttgggatgg
000 xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx gcctcaggaa acgccaggcc
601 tacaggtgga gttggagctg tggctctgct aattgggcca aatgctcctg taatttttga
661 ccgagggctt cgtgggacac acatgcagca tgcctacgac ttttacaagc ctgacatgct
721 ctctgaatac cctgtggtag atggaaaact ctccatacag tgctacctca gcgcattgga
781 ccgctgctat tctgtctacc gcaaaaagat ccgggcccag tggcagaaag agggaaagga
000 xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx catgatcttt cactcgccat actgtaaact
901 ggtgcagaaa tctctagcta ggatgttcct gaatgacttt cttaacgatc aaaacagaga
000 xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx gtgaaattag aagatactta
1021 cttcgacaga gatgtggaaa aggcatttat gaaggctagt gctgagctat tcaaccagaa
1081 aacaaaggca tctttgcttg tatcgaatca aaatggaaac atgtacacat cctctgtata
1141 cggttccctt gcttctgttc tggcacagta ctcacctcaa cagttggccg ggaagaggat
1201 tggagtgttc tcttacggtt ctggcttggc tgccacactc tactccctta aagtcacaca
1261 agatgccaca ccaggatctg ctcttgacaa aataacagca agtttatgtg accttaagtc
1321 aaggcttgac tcaagaacgt gtgtggcacc agacgtcttt gctgaaaaca tgaagctcag
1381 agaggacaca catcacttag ccaactatat tccccagtgt tcaatagatt cactcttcga
1441 aggaacatgg tatctagtca gagtggatga aaagcacaga agaacttacg cccggcgtcc
1501 ctccacaaat gaccacagtt tggatgaagg agtgggactt gtgcattcaa acacagctac
1561 agagcatatt ccaagcccgg ctaagaaagt gccaagactt cctgcgacct cgggcgaacc
13
1621 tgagtcggct gtcatcagta acggggagca ctgagagccg tggccttcac agaggctcgg
1681 ggctggatgg ggtatgggaa acgttagagg aatggatgtc ttgggacaat tttacagatt
1741 acgtgttgct taaaaatgta atgtaactga cacagagccc agaaaactgt tgtgtttttg
1801 gagaagtctc gctgagctcc taacacactt cctgctgtgg gctggccaat ggtgaatgta
1861 ctgcgatggt gttaagggct ctgcagaacg tcatacctcg ctgcatgttt acacgcatgc
1921 gggttaggct tcaaactcgg tctgaactga gtgcttctga ctgcaaaggc agaggtactg
1981 ctgtccagtt taaaaaattg tttttttttt ttttaatgtg taagaatttt tatacttaaa
2041 taaaaaaaag tacctgtagc ttttggggga aaaaaaaaaa cctttttcta ggttggggat
2101 tgtggaattt aaatgttaca cataaactct gcttaatggc aaggcaaaca tttatctttt
2161 tcgaagattt ataaatcctg aagagaaaaa aagagggtat ggttctagga tctggatgaa
2221 ccatcagtga gaaaggttag tcataatcaa gtgagcagaa ggatgctggc gttgagcagg
2281 cctctgtcac agcaaccagg gctctgtggg cacagctgag ggaactttct ggccaggtgc
2341 ccgtgactgc tgctcagctg cactgagatg cagtggagct gctgcacgga agcttgctgt
2401 ggtgctgaac gccttacctg cggataaagt gtaaagtagg agggatgggc agggcactat
2461 taggttacag tgttacagac ccaggttata gacttgacag ctcaaactca ccagacacct
0000 xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx ttttttttta tattgttcaa
2581 tttaaaaaat ttagaaaatt ttaaccttac gttttcacat agtgtgatta gccaaaagga
2641 atttcacttc aagatctaga aatagaattc ataacatttt ttcctaaact ttgactttta
2701 aaacaacgaa aattaccaca tgagatgaac aagaaaattc attagaaagt tctctgggtg
2761 atttttggtg ctgaactgac atgagcctca tagactgtaa aacagaggta gttgaaacta
0000 xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx tgcatttaat tctgtgaagt ttcagttatc
2881 taaaataaac acataaacgt gtaatgtttc agattgcaag gtgagatgta atgtagcatt
2941 tgtaagatat tcttgtcaat attaactggt aggattttga tttgtacagt tttaattggt
3001 taaaatgatc tcattttaac atccactgct atagatgaat gatgtaagct cagatttaat
3061 gaatggtggg gaaatggtgc atgtaatttt ttcgcaagta tcgagagttc tgtatgtttt
3121 gaaaagaata atttaacgtt tgggttgcca ggaagggggc tttcccagag tccattgcca
3181 ggcgttgggc aagcctcgca atgttggcac ggagcgttaa ccacacctta ctaatagcaa
3241 ggggaataac tttgaaataa agttttagac aaata
Company will provide cellular material, known to contain mRNA targets of
interest, as necessary for refinement and optimization of the assays. If
available, Company will provide Institution with 1) cDNA clones for the above
three gene expression assay targets for use as controls, 2) information about
any closely related mRNA species that might be expected to exhibit cross
reactivity. Following evaluation of the Invader assays, Company may share with
Institution certain information about relevant mRNA levels of these genes as
determined by the Company or a third party using other mRNA quantitation
technologies.
Development of Two (2) Polymorphism and Mutation Assays
Company has identified two single nucleotide polymorphisms which Institution
will develop Polymorphism and Mutation detection assays as specified below.
1) [****]
For reference, the sequence from nt3301 to 3720, with the wt allele of the
polymorphic site shown large and bolded, is included below:
cctgggcaag aagtcgctgg agcagtgggt gaccgaggag gccgcctgcc tttgtgccgc cttcgccaac
cactccggtg ggtgatgggc agaagggcac aaagcgggaa ctgggaaggc gggggacggg gaaggcgacc
ccttacccgc atctcccacc cccaggacgc ccctttcgcc ccaacggtct cttggacaaa gccgtgagca
acgtgatcgc ctccctcacc tgcgggcgcc gcttcgagta cgacgaccct cgcttcctca ggctgctgga
cctagctcag gagggactgaaggaggagtc gggctttctg cgcgaggtgc ggagcgagag accgaggagt
ctctgcaggg cgagctcccg agaggtgccg gggctggact ggggcctcgg aagagcagga tttgcataga
14
[****]
2) [****] The polymorphism changes the NAT1 poly adenylation signal from taataaa
to taaaaaa.
Company can provide 20 human genomic DNA samples for evaluation. Company does
not currently have an assay for this polymorphism, and cannot develop a
high-throughput Taqman assay for this SNP due to high AT content in the region
and the presence of a 9 unit trinucleotide repeat just upstream of the
polyadenylation signal. Based on the literature available Company expects to
detect a reasonably high frequency of heterozygotes at the targeted locus.
Company may be able to provide additional genomic DNA samples for evaluation,
if deemed necessary.
EVALUATION OF P450 EXPRESSION INVADER ASSAYS
In order to test Institution's assays for quantitating P450 induction, Company
will provide 24 blind-coded samples of rat liver RNA. Company can provide total
XXX, xXXX, xx both and non-proprietary information about such samples such as
tissue source, time of induction and inducing agent if deemed necessary or
desirable by Institution or Company. Samples will include negative controls and
positive controls for induction of several rat liver cytochrome P450 isoforms.
Ideally, Company would like all 24 samples to be analyzed for expression of ALL
rat cytochromes P450 for which Institution has developed assays; at the minimum,
Company needs data on the 2B and 3A families.
