# US 7,407,750 B2 · Marker assisted selection of bovine for improved milk composition

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- Number: US 7,407,750 B2
- Document ID: US7407750B2
- Status: Granted
- Kind: B2
- Application number: 10/473,683
- Filed: 2002-08-16
- Published: 2004-12-16
- Granted: 2008-08-05
- Earliest claimed: 2002-06-05
- Inventors: Sarah Blott; Jong-Joo Kim; Anne Schmidt-Kuntzel; Anne Cornet; Paulette Berzi; Nadine Cambisano; Bernard Grisart; Latifa Karim; Patricia Simon; Michel Georges; Frederic Farnir; Wouter Coppieters; Sirja Moisio; Johanna Vilkki; Dave Johnson; Richard Spelman; Christine Ford; Russell Snell
- Web page: https://patentgrep.com/patent/US7407750B2

## CPC

- C12Q 1/6888: for detection or identification of organisms
- C12Q 2600/156: Polymorphic or mutational markers
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present invention provides a method of genotyping bovine for improved milk production traits by determining the GHR genotypic state of said bovine, wherein the GHR gene and polymorphisms within said gene have been found to be associated with such improved milk production traits.

## Claims

### Claim 1

A method of selecting a bovine, with a genotype indicative of at least one altered milk production trait, the method comprising the steps:
a) identifying a bovine with thymidine at polymorphic nucleotide position 658 in the growth hormone receptor (GHR) gene of SEQ ID NO: 2, wherein the codon including the thymidine at nucleotide position 658 of SEQ ID NO: 2 encodes phenylalanine at amino acid position 279 in SEQ ID NO: 5; and correlating the presence of the thymidine at position 658 of SEQ ID NO: 2 with at least one altered milk production trait selected from: i) increased milk volume; ii) decreased percentage of milk protein; and iii) decreased percentage of milk fat; and
b) selecting the bovine on the basis of the identification and correlation in a).

### Claim 2

A method of selecting a bovine, with a genotype indicative of at least one altered milk production trait, the method comprising the steps:
a) identifying a bovine with adenosine at polymorphic nucleotide position 658 in the growth hormone receptor (GHR) gene of SEQ ID NO: 2, wherein the codon including the adenosine at nucleotide position 658 of SEQ ID NO: 2 encodes tyrosine at amino acid position 279 in SEQ ID NO: 5; and correlating the presence of the adenosine at position 658 of SEQ ID NO: 2 with at least one altered milk production trait selected from: i) decreased milk volume; ii) increased percentage of milk protein; and iii) increased percentage of milk fat; and
b) selecting the bovine on the basis of the identification and correlation in a).

### Claim 3 (depends on 1)

The method of claim 1, wherein the identification is made by detecting, in a nucleic acid derived from the bovine the presence of thymidine at polymorphic nucleotide position 658 in the GHR gene of SEQ ID NO: 2.

### Claim 4 (depends on 2)

The method of claim 2, wherein the identification is made by detecting, in a nucleic acid derived from the bovine the presence of adenosine at polymorphic nucleotide position 658 in the GHR gene of SEQ ID NO: 2.

### Claim 5 (depends on 3, 4)

The method of claim 3 or 4, wherein detection is made via amplification of a nucleic acid sequence comprising the polymorphic nucleotide position 658 in the growth hormone receptor (GHR) gene of SEQ ID NO: 2.

### Claim 6 (depends on 5)

The method of claim 5, wherein primers consisting of SEQ ID NOs: 6 and 7 are used in the amplification.

### Claim 7 (depends on 3)

The method of claim 3, wherein detection is made using an oligonucleotide ligation assay (OLA).

### Claim 8 (depends on 7)

The method of claim 7, wherein the OLA is performed using a primer comprising the sequence of any one of SEQ ID NOs: 13 or 14.

### Claim 9 (depends on 4)

The method of claim 4, wherein detection is made using an oligonucleotide ligation assay (OLA).

### Claim 10 (depends on 9)

The method of claim 9, wherein the OLA is performed using a primer comprising the sequence of anyone of SEQ ID NOs: 12 or 14.

### Claim 11 (depends on 3, 4)

The method of claim 3 or 4, further comprising the step of hybridizing a probe to the nucleic acid derived from the bovine, wherein the probe:
a) comprises at least 5 contiguous nucleotides of positions 599-702 of SEQ ID NO: 2, wherein said 5 contiguous nucleotides comprises either thymidine or adenosine at the position corresponding to the polymorphic nucleotide position 658 in the GHR gene of SEQ ID NO: 2, and
b) is sufficiently complementary with said nucleic acid so as to bind thereto in 6× sodium citrate/sodium chloride (SSC) at 45° C.;
wherein binding of the probe to the nucleic acid derived from the bovine indicates the presence of specific nucleotide content at the polymorphic nucleotide position 658 of SEQ ID NO: 2.