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CN-118345179-B - PRKAA1 gene as sheep feed conversion rate related molecular marker and application thereof

CN118345179BCN 118345179 BCN118345179 BCN 118345179BCN-118345179-B

Abstract

The invention provides PRKAA genes as molecular markers related to sheep feed conversion rate and application thereof, wherein the molecular markers design primers according to PRKAA gene sequences, DNA is extracted from sheep blood, a C/T polymorphic site exists at 165 th position of an amplified fragment through PCR amplification, DNA sequencing and sequence analysis, then KASPar primers are further used for detecting the polymorphic site of 935 Hu sheep and establishing a least square model, the correlation analysis of genotypes and the feed conversion rate is carried out, and finally, the amplified PRKAA gene fragment can be used as the molecular markers related to the sheep feed conversion rate. The molecular marker can be used for cultivating grain-saving high-quality mutton sheep, provides a genetic engineering means for genetic improvement of sheep feed conversion rate, and has great practical application value.

Inventors

  • GAO FEI
  • TIAN HUIBIN
  • ZHAO YUAN
  • Lian Jiangting
  • LI FADI
  • WANG WEIMIN
  • LI CHENGHAI
  • Pu Mengru
  • WANG LIZHONG
  • LIU JINRUI
  • ZHAO JIAHUI
  • KANG CAIYAN
  • ZHANG DEYIN

Assignees

  • 金川集团股份有限公司

Dates

Publication Date
20260508
Application Date
20240528

Claims (9)

  1. 1. A sheep feed conversion rate related molecular marker is characterized in that the nucleotide sequence of the molecular marker is shown as SEQ ID NO.1, Y at 165 th site of the molecular marker represents C or T, and the C/T mutation at 165 th site of the molecular marker is caused by the fact that the sequence has a C/T mutation at the 165 th site of the molecular marker, so that the C/T polymorphism of sheep PRKAA1 gene at the site is caused.
  2. 2. A PCR primer pair for detecting the molecular marker of claim 1, which comprises a forward primer and a reverse primer, wherein the sequence of the forward primer is shown as SEQ ID NO.2, and the sequence of the reverse primer is shown as SEQ ID NO. 3.
  3. 3. A KASPar primer pair for detecting the molecular marker as claimed in claim 1, which comprises a forward primer A1, a forward primer A2 and a universal reverse primer C, wherein the nucleotide sequence of the forward primer A1 for detecting AlleleA is shown as SEQ ID NO.4, the nucleotide sequence of the forward primer A2 for detecting AlleleG is shown as SEQ ID NO.5, and the nucleotide sequence of the universal reverse primer C is shown as SEQ ID NO. 6.
  4. 4. A kit for detecting the molecular marker according to claim 1, wherein the kit comprises the PCR primer pair according to claim 2 or the KASPar primer pair according to claim 3.
  5. 5. A method of detecting a molecular marker according to claim 1, comprising the steps of: a) Amplifying sheep genomic DNA using the primer pair of claim 2 or 3, or using the kit of claim 4; b) Identifying the polymorphic site of the amplification product obtained in step a).
  6. 6. The method according to claim 5, wherein the polymorphic site is identified by direct sequencing, fluorescent probe, gene chip, or high resolution dissolution profile.
  7. 7. The method according to claim 5, wherein the KASPar primer set according to claim 3 is used for PCR amplification, and after amplification, a BMG PHERAstar instrument is used to detect fluorescent signals and to check the typing result.
  8. 8. Use of a PCR primer pair according to claim 2, a KASPar primer pair according to claim 3 or a kit according to claim 4, or a method according to any one of claims 5-7, in sheep feed conversion detection.
  9. 9. Use of a primer pair according to claim 2 or 3, or a kit according to claim 4, or a method according to any one of claims 5 to 7 in sheep breeding, wherein the breeding is breeding of grain-saving sheep.

Description

PRKAA1 gene as sheep feed conversion rate related molecular marker and application thereof Technical Field The invention belongs to the field of molecular marker preparation, and particularly relates to PRKAA gene serving as a molecular marker related to sheep feed conversion rate and application thereof. Background Mutton is one of the main meat food materials in the current society, and improving the economic benefit of mutton has become one of the most important tasks. In sheep production, the cost of feed is relatively high, so that the improvement of the feed conversion rate of sheep has important research significance. The index designed by genetic improvement of feed investment is reduced on the premise of not influencing the normal growth of animals, and the feed utilization rate of livestock and poultry is reduced. The feed utilization rate of animals refers to the utilization rate of the fed feed, and is mainly influenced by two factors of diet and animals. The feed efficiency (FEED EFFICIENCY, FE) is the short term of the feed conversion rate (Feedconversion ration, FCR), also called feed return, and the feed conversion rate generally refers to the amount of feed consumed by animals with weight gain of 1kg, namely the feed weight ratio (FEED INTAKE/gain, F/G), and is an important economic index for measuring the feed utilization rate for long time. In addition, the weight gain feed ratio (gain/feedintake, G/F) is an index (Lancaster P A,Carstens G E,Jr C D,et al.Phenotypicand genetic relationships of residual feed intake with performance and ultrasound carcass traits in Brangus heifers.Journal of Animal Science,2009,87(12)：3887-3896), for representing the relationship between livestock and poultry weight gain and daily ration feed intake, the feed conversion rate is widely applied at home and abroad, the feed ratio is used in meat product production, and the feed ratio is used in poultry egg production. The (AggreySE,Karnuah A B,Sebastian B,et al.Genetic properties of feed efficiency parameters in meat-typechickens.Genetics Selection Evolution,2010,42(1):1-5.AggreyS E,Rekaya R.Dissection of Koch's residual feed intake:implications for selection.PoultryScience,2013,92(92):2600-2605). researches on improvement of the feed conversion rate start from the aspects of increasing the weight gain of livestock and poultry or the yield of meat and eggs and reducing the feed consumption show that the genetic transmission of the feed conversion rate is 0.26-0.41, belongs to medium genetic traits, is controlled by heredity, can select and match sheep flocks according to scientific data through selecting improvement (Willems O W,MillerS P,WoodB J.Assessment of residual body weight gain and residual intake and body weight gain as feed efficiency traits in the turkey(Meleagris gallopavo).GeneticsSelection Evolution,2013,45(1):1-8)., and is one of the feasible methods (Mount Tao, research on the production performance and the body composition of different RFI fattening lambs and digestion metabolism, gansu agricultural university, 2016). How to determine the scientific basis is one of the main problems to be solved. Currently, most of the indicators are analyzed based on animal phenotypes, with few systematic analysis of sheep feed conversion rates at the genetic level. PRKAA1 (Protein KINASE AMP-ACTIVATED CATALYTIC subnit Alpha 1) is a catalytic Subunit of AMP-activated Protein kinase (AMPK), which plays a key role in （Krishan,S.,D.R. Richardson,andS. Sahni,AMP kinase (<em>PRKAA1</em>). Journal of Clinical Pathology,2014. 67(9): p.758-763.）. in regulating cell energy metabolism through phosphorylation, and a plurality of researches show that PRKAA1 gene is highly expressed in some gastric cancer cells to promote gastric cancer cell proliferation, and pre-clinical data of inhibiting apoptosis (Zhang,Y.,et al.,PRKAA1Promotes Proliferation and Inhibits Apoptosis of Gastric Cancer Cells ThroughActivating JNK1 and Akt Pathways. Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics,2020. 28(3): p.213-223.);PRKAA1 show that PRKAA1 plays a main role in tumor marrow suppression cell (MDSC) induced immunosuppressive activity, overcomes MDSC-driven tumor T cell dysfunction, further improves the effectiveness of immunotherapy （Jimena Trillo-Tinoco,Rosa A. Sierra,Eslam Mohamed,YuCao,Álvaro de Mingo-Pulido,Danielle L. Gilvary,Carmen M. Anadon,Tara Lee Costich,Sheng Wei,Elsa R. Flores,Brian Ruffell,José R. Conejo-Garcia,PauloC. Rodriguez; AMPK Alpha-1 Intrinsically Regulates the Function and Differentiation of Tumor Myeloid-Derived Suppressor Cells.Cancer Res1 October2019;79 (19): 5034–5047.）. clinically, reports that PRKAA1 shows dynamic change （Lee,S.J.,Kang,B.W.,Chae,Y.S. et al. Genetic Variations in STK11,PRKAA1,and TSC1 Associated with Prognosis for Patients with Colorectal Cancer. Ann Surg Oncol 21 (Suppl 4),634–639 (2014). https://doi.org/10.1245/s10434-014-3729-z）.PRKAA1 gene more in tumor and cancer