CN-115820878-B - Transcription factor molecular marker based on regulation and control of fat metabolism of avian myoblasts and application thereof

Abstract

The invention discloses a transcription factor molecular marker based on regulation and control of fat metabolism of avian myoblasts and application thereof, wherein the transcription factor is RXRA gene, the molecular marker is positioned on a chromosome where the RXRA gene is positioned, and comprises a C5952908T mutation site of the chromosome and a nucleotide sequence consisting of upstream and downstream bases of the mutation site, and the mutation site is linked with a C5952944T, A5952945G mutation site and a T5952953C mutation site of a coding region of the chromosome. The invention can judge the individual with high feed efficiency of the poultry by molecular marker screening in early stage through identifying the genotype of the poultry in the genome based on the transcription factor molecular marker for regulating and controlling the fat metabolism of the myoblast of the poultry, provides a direct technical means for the feed efficiency breeding of the poultry, reduces the feeding cost through early breeding, simultaneously genetically improves the feed conversion rate and quickens the genetic progress of breeding.

Inventors

• CHEN XINGYONG
• GENG ZHAOYU
• PAN ZIYI

Assignees

• 安徽农业大学

Dates

Publication Date

20260508

Application Date

20221128

Claims (8)

1. 1. A transcription factor molecular marker based on regulation and control of the fat metabolism of avian myoblasts is RXRA gene, and is characterized in that the molecular marker is a nucleotide sequence containing a sequence shown as SEQ ID NO.1, the molecular marker is positioned on a chromosome where the RXRA gene is positioned and comprises a C5952908T mutation site of the chromosome, and the avian is a meat duck.
2. 2. The molecular marker of claim 1, wherein the mutation site C5952908T of the molecular marker is linked to the C5952944T, A5952945G, T5952953C mutation site of the subsequent chromosomal coding region.
3. 3. Use of a molecular marker according to any one of claims 1-2 for identifying a trait of conversion efficiency in poultry feed.
4. 4. The use of a molecular marker according to claim 3 for identifying the trait of conversion efficiency of poultry feed, wherein if the molecular marker is of T-type, it indicates that the conversion efficiency of poultry feed is high, and if the molecular marker is of C-type, it indicates that the conversion efficiency of poultry feed is general.
5. 5. A method for identifying a trait of conversion efficiency of poultry feed using a molecular marker according to any one of claims 1-2, comprising the steps of: (1) Collecting poultry wing vein blood, and extracting total DNA; (2) Designing a specific amplification primer by taking the molecular marker as a target fragment, and carrying out PCR (polymerase chain reaction) amplification by taking total DNA as a template and utilizing the specific amplification primer to obtain an amplification product; (3) Judging the molecular marker type of the amplified product, if the molecular marker type is T-type, the poultry feed conversion efficiency is high, and if the molecular marker type is C-type, the poultry feed conversion efficiency is general.
6. 6. The method for identifying a trait for conversion efficiency of poultry feed according to claim 5, wherein the method for judging the molecular marker type of the amplified product in step (3) is as follows: If the genotype corresponding to the molecular marker type is TT homozygosity, the poultry feed conversion efficiency is high; If the genotype corresponding to the molecular marker type is CT heterozygous, the poultry feed conversion efficiency is higher; If the genotype corresponding to the molecular marker type is CC homozygous, the poultry feed conversion efficiency is general.
7. 7. The method for identifying the feed conversion efficiency trait of poultry according to claim 6, wherein the method for judging the molecular marker type of the amplified product comprises analyzing the site polymorphism and distribution of the RXRA gene coding region where the amplified product is located by PCR amplification and enzyme digestion sequencing, and determining the feed conversion efficiency of an individual poultry according to the haplotype by dividing the haplotype according to the linkage relation of the polymorphic sites: if the haplotype is TTTTGGCC, the poultry feed conversion efficiency is high; if the haplotype is TCTCGATC, TCTCGGCC, the poultry feed conversion efficiency is higher; if the haplotype is CCCCAATT, the conversion efficiency of the poultry feed is general.
8. 8. The method of identifying an avian feed conversion efficiency trait according to claim 5 wherein the sequence of the specific amplification primer is: F: GTCAGACCTGAGGGCACAA R: CAATTCTGCTCACCGCAA。

Description

Transcription factor molecular marker based on regulation and control of fat metabolism of avian myoblasts and application thereof Technical Field The invention relates to the technical field of molecular markers, in particular to a transcription factor molecular marker based on regulation and control of fat metabolism of poultry myoblasts and application thereof. Background The cost of the feed is the largest cost in the poultry cultivation process, and the cost is about 50-70% of the cultivation cost. Since there is a certain competition relationship between humans and poultry for grains, and with the increasing cost of feed, feed consumption becomes the factor that affects poultry farming the greatest. Therefore, how to improve the efficiency of poultry farming feed and avoid the waste of feed energy becomes one of the core tasks of poultry breeding work in recent years. Fat metabolism is a major factor in the variation of feed efficiency. Therefore, how to effectively block excessive fat deposition in cells, avoid energy consumption due to fat metabolism and increase feed consumption becomes an effective way for improving the feed efficiency of poultry. Nuclear Receptors (NR) control tissue development, homeostasis and metabolism primarily as transcriptional regulators. Retinoic acid X receptor Alpha (RXRA) is one of the nuclear receptor superfamily members and is mainly involved in lipid metabolism, cell differentiation, glucose and energy metabolism. It has been found that β -hydroxybutyrate inhibits fatty acid oxidation and ketone body formation by inhibiting sheep RXRA signals. Construction LNCRNADANCR binds to RXRA to enhance PI3K/AKT signaling and breast carcinogenesis by modulating gsk3β to increase its serine 49/78 phosphorylation. miR-27a enhances sheep preadipocyte proliferation and inhibits differentiation thereof by targeted inhibition of RXRA expression. RXRA promotes the transcription of PLIN1 gene in PPARG independent mode during the differentiation of chicken precursor fat, and further promotes the differentiation of chicken precursor fat cells. However, at present, no report is made as to whether RXRA would have an effect on poultry feed utilization, and how it would have an effect. Disclosure of Invention The invention aims to provide a transcription factor (RXRA) molecular marker for regulating and controlling the fat metabolism of poultry myoblasts and application thereof, wherein the expression quantity of RXRA in the myoblasts is directly related to the fat metabolism process in the cells, the activity of RXRA in the cells is directly influenced by the lipid metabolism in the cells, the invention analyzes the influence of RXRA on the utilization efficiency of poultry feed, the method researches amino acid changes and protein active site conformational changes caused by linkage combination of a plurality of polymorphic sites in RXRA coding region, so as to obtain influencing factors for expression regulation of downstream fat anabolism genes, and develops a molecular marker which is closely linked with the utilization efficiency of poultry feed, and can judge poultry high-feed-efficiency individuals in early stage through molecular marker screening by identifying the type of the molecular marker. The invention realizes the above purpose through the following technical scheme: A transcription factor molecular marker based on regulation and control of the fat metabolism of avian myoblasts is RXRA gene, and the molecular marker is positioned on a chromosome where the RXRA gene is positioned, and comprises a C5952908T mutation site of the chromosome and a nucleotide sequence consisting of upstream and downstream bases of the mutation site, wherein the mutation site is linked with a C5952944T, A5952945G mutation site and a T5952953C mutation site of a coding region of the chromosome. Further, the molecular marker is a nucleotide sequence containing a sequence shown as SEQ ID NO.1 or a mutant sequence thereof, wherein the mutant sequence refers to a C5952908T, C5952944T, A5952945G, T5952953C mutation site of a chromosome coding region corresponding to C103T, C139T, A140G, T148C of SEQ ID NO.1, and one or more mutations are generated in the C103T, C139T, A140G, T C. The invention also provides application of the molecular marker in identifying the transformation efficiency character of poultry feed. Furthermore, if the molecular marker is of the C type, the poultry feed conversion efficiency is high, and if the molecular marker is of the T type, the poultry feed conversion efficiency is general. The invention also provides a method for identifying the transformation efficiency character of the poultry feed by using the molecular marker, which comprises the following steps: (1) Collecting poultry wing vein blood, and extracting total DNA; (2) Designing a specific amplification primer by taking the molecular marker as a target fragment, and carrying out PCR (polymerase chain reaction) amplif