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Swine AMPKγ3R200Q mutation PCR test

avian & livestock genetic test data sheet

Swine AMPKγ3R200Q mutation (RN-) detection by PCR + RFLP

Test code GS003 - Identification of AMPKγ3R200Q mutation, also know as "RN-" in pigs, by PCR+RFLP

The presence of a dominant mutation (sometimes denoted RN-) in Hampshire pigs which significantly affects meat quality and processing yield was first recognized by segregation analysis of phenotypic data. Meat from RN pigs has a low ultimate pH (measured 24 hours after slaughter), a reduced water-holding capacity, and gives a reduced yield of cured cooked ham. These characteristics are due to a ~70% increase in muscle glycogen content in animals that are homozygous or heterozygous carriers of this mutation (Scheffler et al., 2016). However, animals with this mutation do not develop any pathological effects, including any glycogen storage diseases.

Through extensive sequencing efforts, the mutation has been characterized to be a non-conservative substitution (R200Q) in the PRKAG3 gene, which encodes a muscle-specific isoform of the regulatory γ subunit of adenosine monophosphate activated protein kinase (AMPK). AMPK is a heterotrimeric serine/threonine kinase composed of a catalytic α, and regulatory β and γ subunits. AMPK is activated by an increase in the ratio of AMP to adenosine triphosphate (AMP:ATP). Activated AMPK turns on ATP-producing pathways and inhibits ATP-consuming pathways. AMPK can also inactivate glycogen synthase, the key regulatory enzyme of glycogen synthesis, by phosphorylation. Several isoforms of the three different AMPK subunits are present in mammals. The R200Q mutation is located in the protein domain that is essential for binding AMP or ATP, and the mutation reduces AMP dependence and elevates basal activity of AMPK.

This mutation has beneficial effects on meat content but detrimental effects on processing yield. Most breeders would like to eliminate the AMPKγ3R200Q mutation from their herds because of its negative effect on processing yield. Genetic screening is crucial to select pigs for future breeding that are clear of the mutation.

Utilities:

  • Identify this mutation in swine
  • Help remove this mutation from herds
  • Enable breeding of pigs with specific genotypic characteristics

References:
Scheffler, T.L., Park, S., Roach, P.J. and Gerrard, D.E. (2016) Gain of function AMP-activated protein kinase γ3 mutation (AMPKγ3R200Q) in pig muscle increases glycogen storage regardless of AMPK activation. Physiol. Rep. 4: e12802.

Specimen requirements: Buccal swab, or 0.2 ml whole blood in EDTA (purple top) tube, or 0.2 ml fresh, frozen or fixed tissue.

Contact Zoologix if advice is needed to determine an appropriate specimen type for a specific diagnostic application. For specimen types not listed here, please contact Zoologix to confirm specimen acceptability and shipping instructions.

For all specimen types, if there will be a delay in shipping, or during very warm weather, refrigerate specimens until shipped and ship with a cold pack unless more stringent shipping requirements are specified. Frozen specimens should be shipped so as to remain frozen in transit. See shipping instructions for more information.

Turnaround time: 3 business days

Methodology: PCR + restriction fragment length polymorphism

Normal range: Not detected

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