EP1673382A1 - Snp de recepteur adrenergique pour ameliorer les caracteristiques de traite - Google Patents

Snp de recepteur adrenergique pour ameliorer les caracteristiques de traite

Info

Publication number
EP1673382A1
EP1673382A1 EP04784592A EP04784592A EP1673382A1 EP 1673382 A1 EP1673382 A1 EP 1673382A1 EP 04784592 A EP04784592 A EP 04784592A EP 04784592 A EP04784592 A EP 04784592A EP 1673382 A1 EP1673382 A1 EP 1673382A1
Authority
EP
European Patent Office
Prior art keywords
dna
milking
allele
beta2
pair
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP04784592A
Other languages
German (de)
English (en)
Other versions
EP1673382A4 (fr
Inventor
Robert J. Collier
Michael Lohius
Michael Grosz
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Monsanto Co
University of Arizona
Original Assignee
Monsanto Co
University of Arizona
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Monsanto Co, University of Arizona filed Critical Monsanto Co
Publication of EP1673382A1 publication Critical patent/EP1673382A1/fr
Publication of EP1673382A4 publication Critical patent/EP1673382A4/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • C12Q1/683—Hybridisation assays for detection of mutation or polymorphism involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/124—Animal traits, i.e. production traits, including athletic performance or the like
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the present invention relates, in general, to the field of molecular biology, human and bovine genetics, and desirable milking characteristics. In particular this invention provides a method for using genetics to predict the milking characteristics of cows.
  • Various publications or patent are referred to throughout this application to describe the state of the art to which the invention pertains. Each of this publications or patents is incorporated by reference herein. Complete citations of scientific publications are set forth in the text or at the end of the specification.
  • ADRB2 bovine beta2-adrenergic receptor
  • a method for breeding cows for improved milking characteristics including the step of screening the genotype of the parents of the cow for the allele associated with a desired SCS phenotype.
  • the method has the steps of obtaining a DNA sample from a bull to be tested for the desired SCS phenotype; and detecting the presence of an adenine at position 11 in a gene encoding a bovine beta2-adrenoreceptor.
  • the method can be performed by direct sequencing, primer extension, restriction length fragment polymorphism, and allele-specific hybridization.
  • a method of identifies a bull whose daughter cows have a short milking duration
  • This method has the steps of obtaining a sample of DNA from a bull; combining the DNA with a pair of PCR probes comprising SEQ IDs 1 and 2 or SEQ IDS 3 and 4; incubating the DNA under conditions permitting the DNA bounded by the PCR probes to produce DNA amplicons; isolating the DNA amplicons; combining the DNA amplicons with a restriction enzyme specific for CCCGGG for a sufficient time to produce a mixture of DNA fragments from the amplicons comprising CCCGGG; applying the DNA fragment mixture to a gel and permitting migration of the mixture components for a time sufficient for them to separate; and observing the sizes of DNA on the gel, with the largest fragments being correlated with the A genotype and with better SCS phenotype, and the smaller fragments being associated with the C genotype and less desirable SCS phenotype.
  • a milking attribute PCR-RFLP kit contains a pair of primers which flank the 11 th nucleotide position of the bovine beta2-adrenoreceptor gene, and a restriction enzyme specific for the CCCGGG site.
  • the restriction enzyme can be Smal.
  • the primer pairs are selected from pair 1 (SEQ ID Nos 1 and 2) or from pair 2 (SEQ ID Nos 3 and
  • ADR2 bovine beta2-adrenoceptor
  • oligonucleotide primers to be used to amplify and perform locus-specific re-sequencing of genomic DNA spanning regions of the bovine ADRB2 gene. These primers were used to amplify genomic DNA from 24 Holstein and 12 Brown Swiss dairy cows. After comparative genomic sequence was obtained, we analyzed the sequence using polyPhred software (Washington University, St. Louis, MO) and identified, among others, a Single Nucleotide Polymorphism (SNP) sequence at position 11, inclusive of the start ATG (hereafter referred to as "Al 1C"). The published sequence indicates the presence of a cytosine (C) nucleotide.
  • SNP Single Nucleotide Polymorphism
  • This polymorphism was found to be associated with milking characteristics.
  • the A allele at the A11C locus is associated with higher somatic cell score (SMS) and therefore with a faster milking speed.
  • SMS somatic cell score
  • the A allele is effective in both the heterozygous and homozygous conditions to improve the performance of the animal.
  • Polymorphism refers generally to the ability of an organism or gene to occur in two or more different forms. In particular for purposes of the present invention, “polymorphism” refers to two or more different forms of the same gene.
  • SNP Single Nucleotide Polymorphism
  • a “Restriction Enzyme” refers to an endonuclease which bins to double stranded
  • Restriction enzymes are denoted by three-letter abbreviations followed by a strain designation and/or a Roman numeral distinguishing different enzymes from the same species or strain. Recognition sequences are written 5' to 3' for one strand only. Examples of restriction enzymes include Smal, BamHi, Bell, EcoRL Hindlll, and Xbal.
  • allelic refers generally to any of one or more alternative forms of a given gene or DNA segment; both or all alleles of a given gene are concerned with the same trait or characteristic, but the product or function coded for by a particular allele differs qualitatively and/or quantitatively from that coded for by other alleles of that gene.
  • allelic pair i.e., the two alleles of a given gene
  • the polynucleotides of the present invention may be prepared by two general methods: (1) They may be synthesized from appropriate nucleotide triphosphates, or (2) they may be isolated from biological sources. Both methods utilize protocols well known in the art. The availability of nucleotide sequence information enables preparation of an isolated nucleic acid molecule of the invention by oligonucleotide synthesis. Synthetic oligonucleotides may be prepared by the phosphoramidite method employed in the Applied Biosystems 37A DNA Synthesizer or similar devices.
  • Buffer RLT (Qiagen)(with 10 ⁇ l jS-mercaptoethanol added per 1 ml buffer) was added to the leukocytes; for a starting blood volume of 4.0 ml or less, 2.0 ml was added; and for a greater blood volume, 4.0 ml was added.
  • the mixture was homogenized until the sample was homogeneous.
  • An equal volume of 70% alcohol was added to the homogenized lysate and vigorously shaken.
  • RNA sample purified as described above were added 0.1 volume of 10X DNase I Buffer and 1 ⁇ l of DNase I (2 units) to the RNA, which solution was mixed gently and incubated at 37°C for 20-30 min.
  • Ambion DNase Inactivation reagent was resuspended by flicking or vortexing. From that tube, the greater of 0.1 volume or 5 ⁇ l was added to the sample and mixed well. The tube was incubated for 2 min at room temperature. The tube was next centrifuged at 10,000 g for about 1 min to pellet the DNase inactivation reagent.
  • RNA concentration was then used to determine RNA concentration, and 1 ⁇ g
  • RNA using Omniscript (Qiagen) for Real Time PCR analysis The Omniscript 10X Buffer RT, dNTP mix and RNase-free water were first thawed and mixed by vortexing. Qiagen RNase inhibitor was diluted to a final concentration of 10 units/ ⁇ l in lx Buffer RT and mixed by vortexing briefly. Master mix (2.0 ⁇ l of lOx Buffer RT, 2.0 ⁇ l dNTP mix, 2.0 ⁇ l oligo dT primer (10 uM), 0.5 ⁇ l Rnase inhibitor (10 units/ ⁇ l), 1.0 ⁇ l Omniscript reverse transcriptase and double distilled water) was prepared on ice. Master solution was distributed to the various tubes, and template RNA (0.6 to 0.68 ⁇ l/tube) was added and mixed. The resulting solutions were incubated at 37°C for 60 min.
  • Bovine DNAs are available from the Cooperative Dairy DNA Repository
  • CDDR CDDR population
  • MS milking speed
  • Table 2 contains average estimated transmitting abilities (ETA) for
  • results from statistical analyses of the data indicate that genotypes AA, AC and CC are significantly different from the mean SCS (p ⁇ 0.025) and that genotypes AA and AC are significantly higher than the mean SCS (p ⁇ 0.01). Furthermore, animals with the CC genotype have significantly different (i.e., lower) SCS phenotypes than animals with AA and AC genotypes (p ⁇ 0.05).
  • Table 3 shows the distribution of milking speed (MS) classifications according to genotype
  • Table 4 illustrates the average ETA for MS by genotype. Because of the small number of AA animals, there is not a significant association between MS and Al lC genotype at the 5% level. However, the trend in Table 3 clearly shows that the CC genotype has a lower proportion of bulls transmitting fast MS.
  • the assay results correlate with the SCS and enable the choice of animals with improved MS and mastitis resistance.
  • these discoveries can also be used to select animals in an attempt to effect population changes in MS and mastitis resistance. Applying the assay will enable the selection of animals with SCS which will ultimately improve the herd averages for MS and mastitis resistance.
  • This invention also includes a kit containing reagents that can be used to identify allelic composition at the loci described herein.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé pour dépister l'allèle qui est associé à un phénotype SCS souhaité, consistant : à obtenir un échantillon d'ADN d'un taureau devant être soumis au dépistage du phénotype SCS souhaité, et ; à détecter la présence d'une adénine au niveau de la position 11 d'un gène codant un bêta2-adrénorécepteur bovin. Cette invention concerne en outre un nécessaire d'attributs de traite PCR-RFLP contenant une paire d'amorces qui s'accolent à côté de la 11ème position nucléotidique du gène codant un bêta2-adrénorécepteur bovin, ainsi qu'une enzyme de restriction qui est spécifique du site CCCGGG, et qui peut être l'enzyme SmaI.
EP04784592A 2003-09-23 2004-09-21 Snp de recepteur adrenergique pour ameliorer les caracteristiques de traite Withdrawn EP1673382A4 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US50511303P 2003-09-23 2003-09-23
PCT/US2004/030774 WO2005030789A1 (fr) 2003-09-23 2004-09-21 Snp de recepteur adrenergique pour ameliorer les caracteristiques de traite

Publications (2)

Publication Number Publication Date
EP1673382A1 true EP1673382A1 (fr) 2006-06-28
EP1673382A4 EP1673382A4 (fr) 2006-10-18

Family

ID=34392977

Family Applications (1)

Application Number Title Priority Date Filing Date
EP04784592A Withdrawn EP1673382A4 (fr) 2003-09-23 2004-09-21 Snp de recepteur adrenergique pour ameliorer les caracteristiques de traite

Country Status (7)

Country Link
EP (1) EP1673382A4 (fr)
CN (1) CN100422202C (fr)
AU (1) AU2004276248A1 (fr)
BR (1) BRPI0414583A (fr)
CA (1) CA2539551A1 (fr)
RU (1) RU2006113558A (fr)
WO (1) WO2005030789A1 (fr)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008140467A2 (fr) * 2006-09-29 2008-11-20 Pfizer Inc. Marqueurs génétiques et procédés permettant d'améliorer les caractères de productivité et d'adaptation chez les animaux laitiers
CN101693923B (zh) * 2009-11-09 2012-02-22 山东奥克斯生物技术有限公司 筛选耐热性奶牛的HSP70AlA基因SNP位点、应用及试剂盒
CN110106250B (zh) * 2019-05-28 2020-11-27 中国农业大学 与奶牛围产期代谢疾病抗性相关的分子标记及应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990010714A1 (fr) * 1989-03-15 1990-09-20 Wisconsin Alumni Research Foundation Substance genetique de marquage de vaches laitieres a production superieure de lait
WO1999019512A1 (fr) * 1997-10-10 1999-04-22 University Of Cincinnati POLYMORPHISMES DU RECEPTEUR β-ADRENERGIQUES

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6593092B2 (en) * 2000-04-04 2003-07-15 Abbott Laboratories Beta 2 adrenergic polymorphism detection

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990010714A1 (fr) * 1989-03-15 1990-09-20 Wisconsin Alumni Research Foundation Substance genetique de marquage de vaches laitieres a production superieure de lait
WO1999019512A1 (fr) * 1997-10-10 1999-04-22 University Of Cincinnati POLYMORPHISMES DU RECEPTEUR β-ADRENERGIQUES

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO2005030789A1 *

Also Published As

Publication number Publication date
BRPI0414583A (pt) 2006-11-07
CN100422202C (zh) 2008-10-01
RU2006113558A (ru) 2007-11-10
EP1673382A4 (fr) 2006-10-18
AU2004276248A1 (en) 2005-04-07
CA2539551A1 (fr) 2005-04-07
CN1852916A (zh) 2006-10-25
WO2005030789A1 (fr) 2005-04-07

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