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Fine mapping of chromosome 17 quantitative trait loci and use of same for marker assisted selection

A marker-assisted selection, quantitative trait technology, applied in the detection of genetic differences, the field of genetic variation, can solve problems such as thermal stability and ligand affinity reduction

Inactive Publication Date: 2006-07-26
IOWA STATE UNIV RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mutations in this region of human TBG result in reduced thermostability and ligand affinity

Method used

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  • Fine mapping of chromosome 17 quantitative trait loci and use of same for marker assisted selection
  • Fine mapping of chromosome 17 quantitative trait loci and use of same for marker assisted selection
  • Fine mapping of chromosome 17 quantitative trait loci and use of same for marker assisted selection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0186] Cathepsin Z (CTSZ) PCR-RFLP Test

[0187] AlwNI polymorphism

[0188] Primer

[0189] CT04F: 5'GGC ATT TGG GGC ATC TGG G 3' (SEQ ID NO: )

[0190] CT04R: 5' ACT GGG GGA TGT GCT GGT T 3' (SEQ ID NO: )

[0191] PCR conditions:

[0192] 10×Promega buffer

[0193] Combine 10 µL of Mixture 1 and DNA in a reaction tube and cover with mineral oil. The following PCR program was run: 94°C for 3 minutes; 35 cycles of 94°C for 30 seconds, 62°C for 30 seconds and 72°C for 30 seconds; followed by a final extension at 72°C for 5 minutes.

[0194] AlwNI digestion reaction

[0195] The buffer, enzyme and water are made into a mixture. Add 5 µL to each reaction tube containing the DNA. Incubate at 37°C for at least 4 hours, preferably digest overnight. 4 μL of loading dye was mixed with 6 μL of digested PCR product and the total volume was loaded on a 3% agarose gel. The desired AlwNI pattern is shown in Figure 2A.

Embodiment 2

[0197] GNAS PCR-RFLP test

[0198] BbsI polymorphism

[0199] A. Primers

[0200] GN03F: 5'AAG CAG GCT GAC TAC GTG 3' (SEQ ID NO: )

[0201] GN03R: 5' TCA CCA CAA GGG CTA CCA 3' (SEQ ID NO: )

[0202] PCR conditions:

[0203] 10×Promega buffer

[0204] Combine 10 µL of Mixture 1 and DNA in a reaction tube and cover with mineral oil. The following PCR program was run: 94°C for 3 minutes; 35 cycles of 94°C for 30 seconds, 60°C for 30 seconds and 72°C for 30 seconds; followed by a final extension at 72°C for 5 minutes.

[0205] BbsI digestion reaction

[0206] The buffer, enzyme and water are made into a mixture. Add 6 µL to each reaction tube containing the DNA. Incubate at 37°C for at least 4 hours, preferably digest overnight. 4 μL of loading dye was mixed with 6 μL of digested PCR product and the total volume was loaded on a 3% agarose gel. The desired BbsI pattern is shown in Figure 2B.

Embodiment 3

[0208] MC3R PCR-RFLP test

[0209] MnlI polymorphism

[0210] Primers:

[0211] Forward: 5'GCC TCC ATC TGC AAC CTC T 3' (SEQ ID NO: )

[0212] Reverse: 5' AGC ATG GCG AAG AAG ATG AC 3' (SEQ ID NO: )

[0213] PCR conditions:

[0214] 10×PCR buffer

[0215]Combine Mixture 1 and DNA in a reaction tube and cover with mineral oil. The following PCR program was run: 3 minutes at 94°C; 36 cycles of 30 seconds at 94°C, 1 minute at 54°C and 30 seconds at 72°C; followed by a final extension at 72°C for 10 minutes.

[0216] 2 [mu]L of the PCR reaction was run on a 1.6% agarose gel to confirm successful amplification and negative controls were cleared. Digestion can be performed using the following procedure:

[0217] PCR product

[0218] Mix PCR product, buffer, enzyme and water. Incubate for at least 4 hours, preferably overnight at 37°C. Digests were mixed with loading dye (2:5) and run on 3% NuSieve agarose gels. The ex...

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Abstract

Disclosed herein is fine mapping of a quantitative trait locus on Chromosome 17 which is associated with meat traits, growth and fatness. The quantitative trait locus correlates with several major effect genes which have phenotypic correlations with animal growth and meat quality which may be used for marker assisted breeding. Specific polymorphic alleles of these genes are disclosed for tests to screen animals to determine those more likely to produce desired traits.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application Serial No. 60 / 473,179, filed May 23, 2003, which is incorporated herein by reference in its entirety. [0003] Funding reference [0004] The work on this invention was funded in part by USDA / CSREES Contract Nos. 2003-31100-06019 and 2002-31100-06019, and the Government may have certain rights in this invention. field of invention [0005] The present invention generally relates to the detection of genetic differences among animals. More particularly, the present invention relates to genetic variations as indicators of heritable phenotypes associated with higher meat quality and growth rate and fat deposition. The present invention also discloses methods and compositions for using specific genetic markers and chromosomal regions associated with variation and selection in genotyping animals. Background of the invention [0006] The researchers found that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12NC12N15/00
CPCC12Q1/6881C12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/172
Inventor 马克斯·F.·罗斯柴尔德安东尼奥·拉莫斯金关束
Owner IOWA STATE UNIV RES FOUND
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