Application of muscle development related gene of humped humped camel

Abstract

The invention provides an application of a bactrian camel muscle development related gene in breeding, the bactrian camel muscle development related gene comprises RAPGEF4, PCDH7 and ACVR1C, the RAPGEF4 and the PCDH7 are used for regulating and controlling muscle cell differentiation, and the ACVR1C is used for regulating and controlling ACVR1C fat cell differentiation. The expression quantity of RAPGEF4 is positively correlated with the differentiation of myoblasts in muscle development, the expression quantity of PCDH7 is negatively correlated with the differentiation of myoblasts in muscle development, and the expression quantity of ACVR1C is positively correlated with the differentiation of adipocytes in muscle development. The muscle development related genes RAPGEF4, PCDH7 and ACVR1C of the bactrian camels found by the invention are disclosed for the first time, and can be applied to precise breeding of the bactrian camels.

Description

Technical Field

[0001] The invention belongs to the field of animal gene technology, and in particular relates to an application of a gene related to muscle development of a Sunite Bactrian camel. Background Art

[0002] Bactrian camels are large even-toed ungulate livestock that live in extremely harsh environments such as arid grasslands, deserts, semi-deserts and Gobi areas. After long-term natural selection and adaptive evolution, they have developed strong resistance to high temperatures, high cold, and salt and alkali. The Sunite Bactrian camel is mainly distributed in Xilin Gol League and Ulanqab in Inner Mongolia. Because of its larger physique than other local Bactrian camels, it has better meat, wool and milk production. Camel meat has coarser muscle fibers, less fat content, more lean meat, and camel milk has small fat globules, which are easily digested and absorbed by the human body. Camel meat and camel milk have both high nutrition and excellent "green food" quality, making the Sunite Bactrian camel one of the important economic livestock species in desert grasslands.

[0003] Muscle production and quality are one of the important economic traits of Sunite Bactrian camels. The longissimus dorsi muscle is the most representative skeletal muscle and is also an ideal material for studying meat production performance. Skeletal muscle is a highly heterogeneous tissue, and its muscle fiber content and intramuscular fat ratio directly determine the meat production and quality of livestock. Therefore, studying the genes related to muscle development in Sunite Bactrian camels is undoubtedly of great significance for the precise breeding of meat-producing Sunite Bactrian camels. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide an application of Sunite Bactrian camel muscle development related genes in view of the deficiencies of the above-mentioned prior art. The application discloses that the Sunite Bactrian camel muscle development related genes include RAPGEF4, PCDH7 and ACVR1C. RAPGEF4 and PCDH7 are used to regulate muscle cell differentiation, and ACVR1C is used to regulate ACVR1C adipocyte differentiation.

[0006] In order to solve the above technical problems, the technical solution adopted by the present invention is: an application of a muscle development-related gene of Sunite Bactrian camel in breeding, characterized in that the muscle development-related gene of Sunite Bactrian camel includes RAPGEF4, PCDH7 and ACVR1C.

[0007] Preferably, RAPGEF4 and PCDH7 are used to regulate muscle cell differentiation, and ACVR1C is used to regulate adipocyte differentiation.

[0008] Preferably, the expression level of RAPGEF4 is positively correlated with the differentiation of myoblasts during muscle development, the expression level of PCDH7 is negatively correlated with the differentiation of myoblasts during muscle development, and the expression level of ACVR1C is positively correlated with the differentiation of adipocytes during muscle development.

[0009] Compared with the prior art, the present invention has the following advantages:

[0010] The invention discloses that the muscle development-related genes of Sunite Bactrian camel include RAPGEF4, PCDH7 and ACVR1C. RAPGEF4 and PCDH7 are used for regulating muscle cell differentiation, and ACVR1C is used for regulating ACVR1C adipocyte differentiation, and can provide technical support for the genetic improvement of Sunite Bactrian camel.

[0011] The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is a picture of RNA in situ hybridization used to verify the expression of candidate genes in the longissimus dorsi muscle tissue of Sunite Bactrian camel.

[0013] Figure 2 These are the identification data of the RAPGEF4, PCDH7, and ACVR1C overexpression and interference vectors of the present invention.

[0014] Figure 3 This is the data of RAPGEF4 on the expression of myogenic marker genes in the present invention.

[0015] Figure 4 This is the data of the PCDH7 of the present invention on the expression level of myogenic marker genes.

[0016] Figure 5 The Oil Red O staining pictures of 3T3-L1 cells at 200× after overexpression and interference of the ACVR1C gene in the present invention at 7 days of adipogenic differentiation; (1) is the control group without any treatment, (2) is the Oil Red O staining after overexpression of the ACVR1C gene, and (3) is the Oil Red O staining after interference of ACVR1C.

[0017] Figure 6 The present invention provides data on the expression level of the adipocyte marker gene PPARγ in 3T3-L1 cells after overexpression and interference of the ACVR1C gene at 0, 3, 5, and 7 days of adipogenic differentiation. DETAILED DESCRIPTION

[0018] Example 1

[0019] By performing whole genome resequencing on blood samples of 35 Sunite Bactrian camels, the variant sites screened out were compared with those of wild Bactrian camels and other domestic Bactrian camels, and ultimately 423 selected genes were screened out. After single-cell sequencing of muscle tissue of Sunite Bactrian camels, genes specifically expressed in each cell cluster, genes significantly expressed at pseudo-sequential fulcrums, and selected genes were jointly analyzed, overlapping genes were screened, and their functions were analyzed on the Ensembl website, and ultimately candidate genes closely related to muscle development were determined.

[0020] The longissimus dorsi muscle of the Sunite Bactrian camel was placed in physiological saline, and after cleaning the blood stains and impurities, it was cut into tissue blocks of about 2 mm and stored in a cryopreservation tube containing cryopreservation solution. After being stored at -80℃ for 12 hours, it was transferred to liquid nitrogen for long-term storage. The tissue was fully ground on ice in 2mL pre-cooled Homogeniza Buffer, and the release of cell nuclei and the background of the suspension were observed under a microscope. The tissue homogenate was filtered through 70μm and 40μm filters, and the filtrate was transferred to a 15mL centrifuge tube. Centrifuged at 4℃, 500g for 5min, the supernatant was discarded, and washed twice with 1mL WashBuffer. The precipitate was resuspended in 1mL Blocking Buffer, and the cell nucleus suspension was observed under a microscope after mixing without impurities, and then counted directly. The cell nucleus suspension was stained with 0.4% trypan blue, and the viability was counted under a microscope. If the viability was greater than 80%, it could enter the library construction stage to obtain the single cell suspension of skeletal muscle of the Sunite Bactrian camel.

[0021] The construction and sequencing of the single-cell library were completed by Shanghai Wancheng Biotechnology Co., Ltd. Then, the results of single-cell and whole-genome resequencing of skeletal muscle of Sunite Bactrian camel were integrated, and 21 overlapping genes were obtained as shown in Table 1. Five genes related to muscle development of Sunite Bactrian camel and 16 genes related to intramuscular fat deposition were screened out. Among them, the muscle development-related genes mainly participated in biological processes such as calcium ion binding, cell adhesion, DNA binding, protein binding, and intracellular signal transduction (Table 2). The fat deposition-related genes mainly participated in biological processes such as protein binding, lipid binding, transmembrane transport of substances, cAMP signaling pathway, cell cycle, and enzyme activity (Table 3).

[0022] Table 1 Genes related to muscle development and fat deposition screened by combining selected genes

[0023]

[0024] Table 2 Biological functions of genes related to muscle development

[0025]

[0026] Table 3 Biological functions of genes related to intramuscular fat deposition

[0027] Gene Biological function PLIN1 Protein binding, lipid binding, protein catabolism, cellular response to cold SLC24A4 Calmodulin binding, calcium channel activity, cAMP signaling pathway regulation CYTIP Protein binding, cell adhesion SLC35F4 Transmembrane transporter activity, transmembrane transport RARRES1 Protein binding, negative regulation of cell population proliferation ATRNL1 cAMP signaling pathway regulation, protein binding ATP5ME Transmembrane transporter activity, ATP biosynthesis process, respiratory chain electron transport ADAMTS9 Extracellular matrix organization, endopeptidase activity AP2A1 Protein kinase binding, intracellular protein trafficking, postsynaptic neurotransmitter receptor internalization

[0028] Among the genes screened for muscle development, RAPGEF4 was discovered for the first time, PCDH7 was rarely reported in livestock muscle tissue development, and ACVR1C was not reported in large ruminants. Gene regulatory pathway analysis showed that RAPGEF4 was involved in postsynaptic tissue regulation and cAMP signaling pathway; PCDH7 was involved in calcium ion binding and cell adhesion; ACVR1C was involved in TGFB signaling and NODAL signaling.

[0029] Therefore, the biological functions of RAPGEF4, PCDH7 and ACVR1C were verified by experiments.

[0030] (1) Tissue localization expression test of RAPGEF4, PCDH7 and ACVR1C:

[0031] S1. Cell freezing: Wash the bottom of the culture dish with DPBS, digest with 1 mL of 0.25% trypsin at 37°C for 2 min, add an equal volume of culture medium to terminate the digestion after the cells shrink and become round, gently blow the bottom of the culture dish with a pipette to make all the cells fall off, transfer the cell suspension into a 1.5 mL centrifuge tube, centrifuge at 1500 rpm for 5 min, discard the supernatant, add 1 mL of cell freezing solution to resuspend the cells and count them, transfer them into a cryopreservation tube, freeze them at -80°C for 12 h, and transfer them into liquid nitrogen for long-term storage;

[0032] The cells include C2C12 cells (purchased from Wuhan Saiweier Biotechnology Co., Ltd.) and 3T3-L1 cells (purchased from Haixing Biotechnology Co., Ltd.);

[0033] S2. Cell recovery: Take out the cryopreserved tubes containing cells from liquid nitrogen and quickly transfer them to a 37°C water bath to thaw. When the cell fluid has melted to a small amount of ice, transfer them to a 1.5 mL centrifuge tube and centrifuge at 1500 rpm for 5 min. Discard the supernatant. The precipitate is the cells. Resuspend the precipitate with 1 mL of culture medium and count with a cell counting plate. A certain amount of the precipitate is transferred to a culture dish and incubated at 37°C, 5% CO 2 Culture in an incubator;

[0034] S3. Cell transfection: transfer cells in good growth condition into 6-well plates and incubate at 37°C and 5% CO 2 When the cell density is close to 70% in the incubator, the cells are infected with a virus suspension with an MOI value of 80. After 24 hours, the virus-containing culture medium is replaced with a culture medium containing puromycin and the cells are cultured for 4 days, and finally replaced with normal cell culture medium;

[0035] The two types of cells (C2C12 cells and 3T3-L1 cells) in S1-S3 were cryopreserved, revived, and transfected;

[0036] S4. Cell differentiation induction

[0037] Myogenic differentiation: C2C12 cells were inoculated at a cell density of 5×104 in 6-well plates for culture. When the cells grew to 70%, they were infected with viruses. After 24 hours, they were selected with optimal puromycin for 4 days and then cultured in medium containing 10% FBS. The medium was changed every 2 days. When the cells covered the bottom of the dish and contact inhibition occurred, the medium was replaced with myogenic induction medium containing 2% horse serum for differentiation induction. The medium was changed every 2 days.

[0038] Adipogenic induction: 3T3-L1 cells were inoculated in 6-well plates at a cell volume of 5×104. When the cells grew to 70%, they were infected with viruses. After 24 hours, they were screened with optimal puromycin for 4 days, and then replaced with culture medium containing 10% FBS. The medium was changed every 2 days. When the cells covered the bottom of the dish and contact inhibition occurred, the culture medium was replaced with adipogenic induction medium for differentiation induction. After 2 days of induction, the induction medium was replaced with adipogenic maintenance medium for culture, and the medium was changed every two days. During the induction period, the cell state should be carefully observed. If the cells are found to shrink and become round, the adipogenic maintenance medium should be replaced in time.

[0039] S5. qRT-PCR

[0040] S501, RNA extraction

[0041] After discarding the culture medium in the culture dish, add cell lysis solution at a ratio of 1mL Trizol per 10cm of the bottom of the dish. After standing at room temperature for 5 minutes, repeatedly blow the bottom of the dish to wash the cells, transfer the liquid to a 1.5mL centrifuge tube, add a certain volume of chloroform at a ratio of chloroform: Trizol equal to 1:5, vortex mix, stand at room temperature for 5 minutes, and centrifuge at 12000rpm for 15 minutes at 4℃; after centrifugation, the liquid is divided into three layers, of which the lower layer is chloroform and other substances, the middle white layer is protein, and the upper clear liquid is RNA. Pipette the upper layer of liquid into a new 1.5mL centrifuge tube, add an equal volume of isopropanol to mix, stand at room temperature for 10 minutes, and centrifuge at 12000rpm for 10 minutes at 4℃; the white precipitate after centrifugation is RNA, discard the supernatant, add 1mL 75% ethanol to wash the precipitate, centrifuge at 12000rpm for 5 minutes at 4℃, discard the supernatant, repeat this step once, and dry the ethanol to obtain RNA precipitate. Add 10μLddH 2 Dissolve the precipitate in 5% CO and take 1 μL to detect RNA concentration.

[0042] S502, RT-PCR

[0043] Dilute the RNA concentration to 400-1000 ng / μL, add 2 μL reverse transcriptase, ddH 2Add 10 μL of O and place the prepared reverse transcription system in a PCR instrument. Follow the reverse transcription process of 37°C, 15 min; 85°C, 15 s; and 4°C to reverse transcribe RNA into cDNA and store it at -20°C.

[0044] S503, Q-PCR

[0045] Use ddH 2 O Dilute the cDNA concentration to 100-200 ng / μL, use the untransfected cell line induced cells as the control, GAPDH as the internal reference, and detect the expression of myogenic and adipogenic markers MYOD, MYOG, and PPARγ at 0, 3, 5, and 7 days of induction, respectively. Each sample was repeated 3 times. Amplification primers, real-time fluorescence quantitative PCR system, etc. are shown in the following table:

[0046] Table 4 Real-time fluorescence quantitative PCR amplification primer sequences

[0047]

[0048] Table 5 Real-time fluorescence quantitative PCR reaction system preparation (20 μL)

[0049] Reagents Volume (μL) cDNA 2.0 Upstream primer 0.8 Downstream primer 0.8 TBGreenII 10 ddH2O 6.4

[0050] Table 6 Real-time fluorescence quantitative PCR reaction system

[0051]

[0052]

[0053] The amplification curve and melting curve were obtained according to the Q-PCR results, and the Ct value of the marker gene was calculated, and then the relative expression of the marker gene was calculated using the 2-△△Ct method. GraphPad Prism 8 was used for significance analysis and plotting.

[0054] (2) Oil Red O staining

[0055] Fat, namely triglyceride (TG), is formed by the dehydration of -OH in the carboxyl group of fatty acids and -H in the hydroxyl group of glycerol, and is neutral. Oil Red O (ORO) is a fat-soluble azo dye, a strong fat-soluble reagent and fat staining agent, which can specifically stain neutral lipids such as triglycerides in cells or tissues, but has weak staining effects on phospholipids and steroids. When cells are immersed in Oil Red O staining solution, Oil Red O dissolves in the lipid droplets in the cells, making them red or orange-red for easy observation. When using, first aspirate the culture medium in the culture dish, wash once with DPBS, add 4% paraformaldehyde fixative to fix the cells for 10 minutes, and then gently rinse twice with DPBS. Add staining wash solution that just covers the cells for 20 seconds, aspirate the wash solution, add an appropriate amount of Oil Red O stain solution, and stain for 20 minutes. After removing the staining solution, add the staining wash solution and let it stand for 30 seconds, then remove the wash solution and wash with DPBS for 20 seconds, add an appropriate amount of DPBS and observe the staining results under a microscope.

[0056] (3) RNA in situ hybridization test

[0057] Freshly isolated longissimus muscle of adult Bactrian camel was rinsed in PBS and fixed in tissue fixative at room temperature for more than 12 hours. After fixation, the tissue block was cut into about 3mm thick in a fume hood, dehydrated in low to high gradient alcohol, and then transparent in xylene, immersed in wax and embedded. The tissue wax block was cut into 4μm thick slices using a slicer, spread and scooped by a slicer, and baked in a 62℃ oven for 2 hours. The slices were placed in environmentally friendly dewaxing transparent liquid I for 15 minutes, environmentally friendly dewaxing transparent liquid II for 15 minutes, anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, 85% alcohol for 5 minutes, 75% alcohol for 5 minutes, and DEPC water for soaking. Citric acid (pH6.0) was used to repair the microwave at medium heat for 7 minutes, stop the fire for 7 minutes, and then turn to medium-low heat for 6 minutes. After natural cooling, the tissue was circled with a tissue pen, digested with proteinase K at 40 degrees for 15 minutes, rinsed with pure water and washed with PBS 3 times, each time for 5 minutes.

[0058] In situ hybridization was performed using probes for candidate genes PCDH7, RAPGEF4 and ACVR1C designed by Servicebio. First, add pre-hybridization solution and incubate at 37℃ for 1h. Discard the pre-hybridization solution, add probe-containing hybridization solution, and hybridize overnight in a 40℃ incubator. Wash off the hybridization solution, wash with 2×SSC at 37℃ for 10min, 1×SSC at 37℃ for 2×5min, and 0.5×SSC at room temperature for 10min. Gently shake dry the slices, add preheated branch probe hybridization solution (60μL), and place horizontally in a humidified box for hybridization at 40℃ for 45min. During this process, add 50mL 2×SSC to the bottom of the humidified box to prevent the slices from drying. Remove the hybridization solution and rinse the slices with 2×SSC, 1×SSC, 0.5×SSC, and 0.1×SSC preheated at 40℃ at 40℃ for 5min. Add signal probe hybridization solution at a dilution ratio of 1:200 and incubate at 42℃ for 3h. Then, the sections were washed with SSC in the following order: 2×SSC, 37°C for 10 min; 1×SSC, 37°C for 2×5 min; 0.5×SSC, 37°C for 10 min. DAPI stain was added to the sections, incubated in the dark for 8 min, and then washed with anti-fluorescence quenching sealing agent to seal the sections. The sections were observed under a fluorescence microscope. Figure 1 , Figure 1 This is a picture of RNA in situ hybridization to verify the expression of candidate genes in the longissimus muscle tissue of Bactrian hunchback in Sunite; the picture shows the RNA in situ hybridization of the longissimus muscle of Bactrian hunchback under a 200× microscope, Merge is the co-staining of genes and cell nuclei, blue DAPI is the cell nucleus staining, and the gene is red fluorescence. The results of RNA in situ hybridization experiments showed that RAPGEF4, PCDH7, and ACVR1C were all expressed on the cell membrane of the longissimus muscle tissue of Bactrian hunchback, and RAPGEF4 was also expressed in the cytoplasm. The in situ hybridization experiment first verified the expression and spatial location of the candidate genes RAPGEF4, PCDH7, and ACVR1C in the tissue, and their biological functions need to be further verified through overexpression and interference experiments.

[0059] Identification of overexpression and interference vectors of RAPGEF4, PCDH7, and ACVR1C

[0060] C2C12 cells and 3T3-L1 cells were transfected with RAPGEF4, PCDH7, ACVR1C overexpression and interference lentiviral vectors and lentiviral empty vectors for 24 hours, C2C12 cells were screened with 7μg / mL puromycin, and 3T3-L1 cells were screened with 4μg / mL puromycin for 4 days. The expression levels of RAPGEF4, PCDH7, and ACVR1C genes were detected by qRT-PCR to identify the viral vectors. The corresponding genes in the RAPGEF4, PCDH7, and ACVR1C overexpression vector groups were significantly highly expressed, the gene expression in the interference group was significantly reduced, and the gene expression in the empty vector control group remained unchanged. Figure 2 As shown, A is the identification of RAPGEF4 lentiviral vector, overexpression-RAPGEF4 is the overexpression group, si-RAPGEF4 is the interference group, B is the identification of PCDH7 lentiviral vector, overexpression-PCDH7 is the overexpression group, si-PCDH7 is the interference group, C is the identification of ACVR1C lentiviral vector, overexpression-ACVR1C is the overexpression group, si-ACVR1C is the interference group. NC is the control group without any treatment, and Control is the empty vector group. (*p<0.05, **p<0.01, ***p<0.005).

[0061] Effects of RAPGEF4 and PCDH7 on myogenic differentiation of C2C12 cells:

[0062] C2C12 cells transfected with empty lentiviral vector, RAPGEF4 overexpression lentiviral vector (overexpression-RAPGEF4), and RAPGEF4 interference (si-RAPGEF4) lentiviral vector were induced into myogenesis. The expression levels of myogenesis marker genes MYOD and MYOG on days 0, 3, 5, and 7 of induction were as follows: Figure 3 As shown in the figure, Figure A shows the expression changes of adipocyte marker gene MYOD at 0, 3, 5, and 7 days of myogenic induction differentiation of C2C12 cells under 200× overexpression and interference of RAPGEF4 gene, and Figure B shows the expression changes of adipocyte marker gene MYOG at 0, 3, 5, and 7 days of myogenic induction differentiation of C2C12 cells under 200× overexpression and interference of RAPGEF4 gene. Control is the empty vector group, overexpression-RAPGEF4 is the overexpression group, and si-RAPGEF4 is the interference group (*p<0.05, **p<0.01, ***p<0.005); the results showed that: with the increase of induction days, the expression of muscle marker genes in the overexpression group gradually increased and was significantly higher than that in the empty vector group, and the expression of marker genes in the interference group was significantly lower than that in the empty vector group, indicating that overexpression of RAPGEF4 can promote myogenic differentiation.

[0063] C2C12 cells transfected with PCDH7 overexpression lentiviral vector (overexpression-PCDH7), PCDH7 interference (si-PCDH7) lentiviral vector and lentiviral empty vector were induced into myogenesis. The expression levels of myogenesis marker genes MYOD and MYOG on days 0, 3, 5 and 7 of induction were as follows: Figure 4As shown in the figure, Figure A shows the expression changes of adipocyte marker gene MYOD at 0, 3, 5, and 7 days of myogenic induction differentiation of C2C12 cells after overexpression and interference of PCDH7 at 200×, and Figure B shows the expression changes of adipocyte marker gene MYOG at 0, 3, 5, and 7 days of myogenic induction differentiation of C2C12 cells after overexpression and interference of PCDH7 at 200×, Control is the empty vector group, overexpression-PCDH7 is the overexpression group, and si-PCDH7 is the interference group (*p<0.05, **p<0.01, ***p<0.005); The results showed that: with the increase of induction days, the expression level of myogenic marker genes in the interference group gradually increased and was significantly higher than that in the empty vector group, and the expression level of marker genes in the overexpression group increased slowly, and was significantly lower than that in the empty vector group on the 7th day of induction, indicating that PCDH7 plays an inhibitory role in myogenic differentiation.

[0064] 4. Effect of ACR1C on adipogenic differentiation of 3T3-L1 cells:

[0065] ACVR1C was interfered and overexpressed in 3T3-L1 cells, and adipogenic induction was performed for 0, 3, 5, and 7 days. Figure 6 , Figure 6 The data are for the expression of adipocyte marker gene PPARγ in 3T3-L1 cells after overexpression and interference of ACVR1C gene at 0, 3, 5, and 7 days of adipogenic differentiation. Control is the empty vector group, overexpression-ACVR1C is the overexpression group, and si-ACVR1C is the interference group (*p<0.05, **p<0.01, ***p<0.005); qRT-PCR showed that the expression of adipogenic marker gene PPARγ in ACVR1C interference group (si-ACVR1C) was significantly higher than that in control group (Control) from the 3rd day of induction, and adipogenic differentiation gradually increased with the increase of induction days; while the expression of adipogenic marker gene in ACVR1C overexpression group (overexpression-ACVR1C) was significantly lower than that in control group (Control), and the gene expression increased slowly during the induction period, indicating that interference with ACVR1C can promote the expression of adipocyte marker gene. Oil red O staining was performed on the cells on the 7th day of induction, and the results were obtained. Figure 5 , where (1) is the control group without any treatment, (2) is the Oil Red O staining after overexpression of ACVR1C gene, and (3) is the Oil Red O staining after ACVR1C interference. The results showed that the overexpression group had fewer adipocytes than the control group, and the lipid droplets were smaller and less, while the interference group had more adipocytes than the control group and more and larger lipid droplets, indicating that interference with ACVR1C can promote the differentiation of 3T3-L1 cells into adipocytes.

[0066] Therefore, the present invention screened candidate genes RAPGEF4, PCDH7 and ACVR1C related to muscle development. RNA in situ hybridization showed that RAPGEF4 was expressed in the cell membrane and cytoplasm of the longissimus dorsi muscle of Bactrian camel, and PCDH7 and ACVR1C were mainly located on the cell membrane of the longissimus dorsi muscle. RAPGEF4 and PCDH7 both play a certain regulatory role in myoblast differentiation. RAPGEF4 can promote myoblast differentiation, while PCDH7 inhibits myoblast differentiation; ACVR1C negatively regulates adipocyte differentiation. The candidate genes RAPGEF4, PCDH7 and ACVR1C can all be used in breeding to select Sunite Bactrian camels with better muscle development.

[0067] The above is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent change made to the above embodiment according to the technical essence of the invention still falls within the protection scope of the technical solution of the present invention.

Claims

1. An application of a gene related to muscle development of Sunite Bactrian camel in breeding, characterized in that: The genes related to muscle development in Sunite Bactrian camel include RAPGEF4, PCDH7 and ACVR1C.

2. The use according to claim 1, characterized in that: RAPGEF4 and PCDH7 are used to regulate muscle cell differentiation, and ACVR1C is used to regulate adipocyte differentiation.

3. The use according to claim 1, characterized in that: The expression level of RAPGEF4 was positively correlated with the differentiation of myoblasts during muscle development, the expression level of PCDH7 was negatively correlated with the differentiation of myoblasts during muscle development, and the expression level of ACVR1C was positively correlated with the differentiation of adipocytes during muscle development.

Images