KR101813054B1 - Method for identifying meat quality trait in Berkshire using MYC gene, SNP marker associated thereof and its primer set - Google Patents
Method for identifying meat quality trait in Berkshire using MYC gene, SNP marker associated thereof and its primer set Download PDFInfo
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Abstract
The present invention relates to a method for confirming meat quality improvement of a Berkshire variety pig using a MYC gene of a Berkshire variety pig, a SNP marker and a primer set associated therewith, and more particularly, to a mutation in a MYC gene of a Berkshire variety, A method for confirming whether or not the meat quality of a Berkshire variety pig is improved, a SNP marker and a primer set related to the method, and further, The present invention provides a selection method of breeds of Berkshire variety pigs using the SNP assay kit and the SNP marker for improving the meat quality of the Berkshire variety pig.
Description
The present invention relates to a method for confirming meat quality improvement of a Berkshire variety pig using a MYC gene of a Berkshire variety pig, a SNP marker and a primer set associated therewith, and more particularly, to a mutation in a MYC gene of a Berkshire variety, A method for confirming whether or not the meat quality of a Berkshire variety pig is improved, a SNP marker and a primer set related to the method, and further, The present invention provides a selection method of breeds of Berkshire variety pigs using the SNP assay kit and the SNP marker for improving the meat quality of the Berkshire variety pig.
The pig industry has focused primarily on improving the productivity associated with the quantity of meat and on the distribution of fresh meat, which has been accompanied by a decline in pork quality (Cameron, 1990, Cliplef and McKay, 1993). In Korea, the pattern of consumption of pork consumes a lot of fresh meat because most of it is grilled and pork. Basically, meat quality consumer awareness is an important selection criterion for meat purchase. However, uneven meat quality reduces consumer confidence and decreases willingness to choose good meat (Jung et al., 2011). Therefore, in order to overcome these problems, the industry has developed a meat quality trait improvement system that satisfies the consumer's demand for the best pork quality (Lee et al., 2012).
Of the various pig varieties, colored varieties are generally known to have higher marbling scores than white varieties. In Japan, the price of pork in Berkshire varieties is 50% higher than that of regular commercial pork. In Korea, Berkshire pork is marketed as premium meat (DO et al., 2012). Preference for black pig meat is partly due to increased consumer demand for high quality pork. However, insufficient supply is now an obstacle to increasing consumption. This rise in consumer expectations is fueling active genetic-based research in Berkshire varieties that ultimately is carried out to improve breeding and management techniques for improved pork productivity and quality.
On the other hand, a porcine MYC (v-myelofibrosarcoma tumor oncogene) gene coding for c-MYC protein is composed of 3 exons and 2,464 base pairs and is located on chromosome 4. The c-MYC protein is considered to be an important transcriptional regulator during cell development and the gene expression of the MYC gene is associated with various physiological processes including hormones, growth factors, cytokines, lymphokines, nutritional status, development, Reported. In addition, its function is associated with muscle formation, muscle growth and fat formation. Our previous study (Oh et al., 2014) showed that non-synonymous SNPs in porcine MYC genes may be associated with some economic traits such as age at 90 kg, for example. However, there has been no prior art studying the relationship between porcine MYC gene polymorphism and meat quality characteristics of pigs in the past.
Thus, the present inventors have confirmed the Berkshire variety-specific single base polymorphism and polymorphism of the C- MYC gene having pork quality characteristics of Berkshire variety to complete the present invention.
It is an object of the present invention to provide a primer set for improving meat quality of a Berkshire variety pig using a MYC gene of a Berkshire variety pig.
It is another object of the present invention to provide a SNP marker for improving meat quality of a breed of a Berkshire variety obtained by amplification through a polymerase chain reaction with the above primer set.
Yet another object of the present invention is to provide a SNP assay kit for meat quality improvement of a Berkshire variety pig comprising the primer set, the PCR reaction mixture and the restriction enzyme HFA I for restriction fragment length polymorphism (RFLP) analysis.
It is another object of the present invention to provide a method for confirming the meat quality improvement of a Berkshire variety pig using the MYC gene of a Berkshire variety pig.
It is another object of the present invention to provide a method of selecting breeds of a Berkshire variety pig using the MYC gene of a Berkshire variety pig.
In order to solve the above-described problems, the present invention provides a primer set for improving meat quality of a Berkshire variety pig using Berkshire variety pig MYC gene, which is characterized by a primer set consisting of the nucleotide sequences of SEQ ID NOS: 2 and 3.
The present invention also provides SNP markers for improving meat quality of a Berkshire variety pig characterized by a G3350C SNP marker obtained by amplification through a polymerase chain reaction with the above primer set.
According to a preferred embodiment of the present invention, the G3350C SNP marker may be generated by base substitution at position 3350 (G? C) shown in SEQ ID NO: 1.
The present invention also provides a SNP assay kit for meat quality improvement of a breeder variety of pigs, comprising the aforementioned primer set, PCR reaction mixture and Hha I as a restriction enzyme for restriction fragment length polymorphism (RFLP) analysis.
(A) amplifying a MYC gene comprising the aforementioned SNP marker from a genomic DNA of a sample isolated from a Berkshire variety pig; (b) treating the MYC gene amplified in the step (a) with a restriction enzyme Hha I ; And (c) comparing the polymorphism (RFLP) of the gene fragments obtained in the step (b) with the MYC gene of a Berkshire variety pig.
According to a preferred embodiment of the present invention, when the gene fragment of step (c) is identified as one fragment having 681 bp, it can be determined that the meat quality of the Berkshire variety pig is improved.
According to another preferred embodiment of the present invention, when the gene fragment of step (c) is identified as one fragment having 681 bp, it is more preferable that the fragment having 480 bp and the fragment having 201 bp are 45 minutes after slaughter The pH value indicating the muscle acidity of 45 minutes is high and the heating loss can be low.
(A) amplifying a MYC gene comprising the aforementioned SNP marker from a genomic DNA of a sample isolated from a Berkshire variety pig; (b) treating the MYC gene amplified in the step (a) with a restriction enzyme Hha I ; And (c) comparing the polymorphism (RFLP) of the gene fragments obtained in step (b) with the MYC gene of a Berkshire variety pig.
According to a preferred embodiment of the present invention, when the gene fragment of the step (c) is identified as a single fragment having 681 bp, it can be selected as an improved breed.
The inventors of the present invention confirmed that mutation of a specific site in the MYC gene plays an important role in meat quality such as pH and heat loss of muscles. And it is considered that the economic value is very high. Therefore, it is expected that mutant areas will be utilized as basic data for the development of molecular genetic technology for improving meat quality of Berkshire variety pigs in the future, and it will be possible to develop molecular breeding technology for improving meat quality through continuous research.
Figure 1 shows the genomic structure and polymorphism of the MYC gene of Berkshire variety pigs and the amplified portion of g.3350 G > C with Hha I < / RTI > restriction enzyme.
Hereinafter, the present invention will be described in more detail.
As described above, the consumption pattern of pork consumption in Korea is dominated by roast and pork, which consumes a large amount of fresh meat. Consumer demand for pork of a colored variety, which is considered to be higher in quality than white pork, is increasing, The production volume is insufficient compared to the demand quantity. As a result, interest in Berkshire variety pigs has increased recently, and technology for improving the productivity and quality of Berkshire variety pork has been demanded.
Therefore, the present inventors sought a solution to the above-mentioned problem by providing a method for confirming the meat quality improvement of a Berkshire variety pig using the MYC gene of a Berkshire variety pig, SNP markers and a primer set related thereto. The method for confirming the meat quality improvement of the Berkshire variety pig of the present invention is characterized by using a mutation in the MYC gene of the Berkshire variety pig having a significant correlation with meat quality such as muscle acid pH (pH 45 min ) and cooking loss And mutation in the MYC gene can be utilized as a molecular marker for improving meat quality, so that it is useful for selecting breeds of breeds of breeds of Berkshire variety improved in meat quality.
The terms used in the present invention are defined as follows.
In the present invention, "SNP marker" refers to a SNP that is an allelic variant of any variant type found in the genome.
In the present invention, a "primer" is a single strand of oligonucleotides, which is hybridized under suitable conditions (presence of four different nucleoside triphosphates and a polymerase such as a DNA or RNA polymerase) Lt; RTI ID = 0.0 > template-directed DNA < / RTI > The suitable length of the primer is determined by the characteristics of the primer to be used, but is usually 15 to 30 bp in length. The primer need not be exactly complementary to the sequence of the template, but should be complementary enough to form a hybrid-complex with the template.
In the present invention, a "meat quality improving" means the genotype of the SNP (GG, GC, and CC) to result, after slaughter affecting the meat 45 minutes of back muscle pH (pH 45min) and the heat loss is significantly changed by a high a pH of 45 min and a low heat loss.
All technical terms used in the present invention are used in the sense that they are generally understood by those of ordinary skill in the relevant field of the present invention unless otherwise defined. Also, preferred methods or samples are described in this specification, but similar or equivalent ones are also included in the scope of the present invention.
The present invention provides a primer set for improving meat quality of a Berkshire variety pig using Berkshire variety pig MYC gene, which is characterized by a primer set consisting of the nucleotide sequences of SEQ ID NOS: 2 and 3.
The primer set can amplify a polymorphic site specific to a Berkshire variety located at intron 2 of the MYC gene shown in FIG.
The present invention also provides SNP markers for improving meat quality of a Berkshire variety pig characterized by a G3350C SNP marker obtained by amplification through a polymerase chain reaction with the above primer set.
The G3350C SNP marker may be generated by base substitution at position 3350 (G? C) shown in SEQ ID NO: 1.
The nucleotide sequence shown in SEQ ID NO: 1 was retrieved from the NCBI database as the DNA sequence of the porcine MYC gene (SEQ ID NO: X97040.1 (Genbank)). The nucleotide sequence 3350 base (G → C) shown in SEQ ID NO: 1 has been discovered by the inventors of the present invention, and no method for screening individuals having improved meat quality using the G3350C SNP marker has been known at all.
The present invention also provides a primer set, PCR reaction mixture, and polymorphism (RFLP: restriction fragment length polymorphism) analysis of the restriction enzyme Hha for I < / RTI > for improving meat quality of a Berkshire variety pig.
The SNP assay kit uses the PCR-RFLP technique. To analyze the expression of various mutations in the MYC gene involved in meat quality, a specific restriction enzyme Hha I. These specific restriction enzymes are those selected by the present inventors on the basis of SNPs that reveal SNPs that cause changes in meat quality characteristics.
In Example 3 of the present invention, specifically, the restriction enzyme Hha which can recognize the G3350C SNP site of the present invention I was identified and genotypes were identified as shown in FIG. For example, when amplified with the primers set forth in SEQ ID NOS: 2 and 3, the genotypes can be divided into GG, GC, and CC, and the G allele genotypes are divided into two fragments (480 bp and 201 bp) And one fragment (681 bp) of the C allele genotype was identified.
(A) amplifying a MYC gene comprising the above-described SNP marker of the present invention from a genomic DNA of a sample isolated from a Berkshire variety pig; (b) amplifying the MYC gene amplified in step (a) with restriction enzyme Hha I; And (c) comparing the polymorphism (RFLP) of the gene fragments obtained in the step (b) with the MYC gene of a Berkshire variety pig.
The genomic DNA of step (a) can be obtained from various sources and can be obtained, for example, from muscles, epidermis, blood, bones and organs, and preferably from muscles or blood.
When the starting material is genomic DNA as described above, the genomic DNA can be isolated according to a conventional method known in the art (see Rogers & Bendich (1994)). In addition, when the starting material is mRNA, total RNA is isolated by a conventional method known in the art (Sambrook, J. et al., Molecular Cloning, A Laboratory Manual, 3rd ed., Cold Spring Chomczynski, P. et al., Anal. Biochem. 162: 156 (1987)). The isolated total RNA is synthesized by cDNA using reverse transcriptase. Since the total RNA is isolated from animal cells, it has a poly-A tail at the end of mRNA. CDNA can be easily synthesized using oligo dT primers and reverse transcriptase using such sequence characteristics.
The amplification of step (a) may be performed without limitation as long as the primer capable of amplifying the G3350C SNP site of the SNP marker of the present invention contained in the MYC gene is used. Preferably, the primer is composed of the nucleotide sequence of SEQ ID NOS: 2 and 3 Can be amplified with a primer set.
The restriction enzyme of step (b) is not particularly limited as long as it is a restriction enzyme capable of recognizing the SNP site. Preferably, the restriction enzyme Hha I is appropriate.
The present invention has developed a search technique using the PCR-RFLP method, which is one of the most efficient methods for searching and diagnosing the SNP region by performing a single PCR reaction and enzyme treatment on the MYC gene including the SNP region.
The comparison of the polymorphism (RFLP) in the step (c) may be performed using the restriction enzyme Hha I is to discriminate the genotypes by grasping the band patterns that appear differently according to the processing of I.
If the gene fragment of step (c) is identified as one fragment having 681 bp, the meat quality of the Berkshire variety pig can be judged to be improved.
Specifically, the meat quality of the Berkshire breed pig can be determined according to the muscle pH (pH 45min), and cooking loss after 45 minutes after the slaughter.
The pH of 45 min is an important economic factor affecting the freshness of meat due to post-metabolism, which is related to the ability of the muscle to retain moisture and has a significant effect on the appearance of meat.
As measured in Example 4, this genotype is suggested to play an influential role in the improvement of pork quality, as the pH 45 min was significantly higher in individuals with the CC genotype than those with the GG and GC genotypes.
Also, in the weight loss of heating, a high heating loss shows a low meat quality, and a high marbling score is reported to have a low heating loss, so that a low heating loss is regarded as an advantageous characteristic of meat quality.
As measured in Example 4, the measured weight loss of the GG, CG and CC genotypes were 16.38 ± 0.63, 15.48 ± 0.84 and 15.50 ± 0.67, respectively (Table 5).
Therefore, when the gene fragment of step (c) is identified as a fragment having 681bp object has a CC genotype, CC genotype exhibits a low heat loss and high pH 45min determined that enhanced the meat quality of the Berkshire breed pig can do.
(A) amplifying a MYC gene comprising the above-described SNP marker of the present invention from a genomic DNA of a sample isolated from a Berkshire variety pig; (b) amplifying the MYC gene amplified in step (a) with restriction enzyme Hha I; And (c) comparing the polymorphism (RFLP) of the gene fragments obtained in step (b) with the MYC gene of a Berkshire variety pig.
If the gene fragment of step (c) is identified as a single fragment having 681 bp, it can be selected as an improved breed.
Hereinafter, the present invention will be described in more detail with reference to the following Examples. However, these drawings and the following embodiments are only illustrative of the contents and scope of the technical idea of the present invention, and the technical scope of the present invention is not limited or changed. It will be apparent to those skilled in the art that various modifications and variations are possible within the scope of the technical idea of the present invention based on these examples.
Disclosed animal and DNA isolation
In order to detect breed-specific variation in the MYC gene, four types of pigs of the purebred pigs in Korea, Berkshire, Duroc, Landrase and Yorkshire, Twenty blood samples from each were collected with a vacuum lavender containing an anticoagulant. Genomic DNA was isolated from the blood sample using the G-DEX TM IIb Genomic DNA Extraction Kit (IntronBio Inc., Korea).
To determine meat quality characteristics, muscle samples were obtained from Berkshire carcasses of 378 sows (138 sows, 240 sows) slaughtered at a slaughterhouse in Namwon, Jeonbuk, Korea. Genomic DNA was extracted from muscle samples using the same kit as above. All procedures were carried out in accordance with the manufacturer's instructions.
primer Production and Polymerase Chain Reaction PCR )
The DNA sequence of the porcine MYC gene was retrieved from the NCBI database (Session Number: X97040.1 (Genbank)). As shown in Table 1, the primer 3 program (http://frodo.wi.mit.edu/primer3/) produces 8 primer sets (P1-P8) for full-length sequencing of the MYC gene by PCR Respectively.
(SEQ ID NO)
(bp)
(° C)
(SEQ ID NO: 4)
5'-CAAAAACAACCAACCCCTCT-3 '
(SEQ ID NO: 5)
(SEQ ID NO: 6)
5'-GGGTAGCAGCTGTTCTGGA-3 '
(SEQ ID NO: 7)
(SEQ ID NO: 8)
5'-CACACTTGGTGAACCACCTC-3 '
(SEQ ID NO: 9)
(SEQ ID NO: 10)
5'-GTAGGAGGCCAGCTTCTCTG-3 '
(SEQ ID NO: 11)
(SEQ ID NO: 12)
5'-AAGGGAGGCAATTCATTCAC-3 '
(SEQ ID NO: 13)
(SEQ ID NO: 14)
5'-ACCCTGGCACAGAGTTATCC-3 '
(SEQ ID NO: 15)
(SEQ ID NO: 16)
5'-ACTGGATCATGCATTCGAGA-3 '
(SEQ ID NO: 17)
(SEQ ID NO: 18)
5'-TTAAGGCCCAGACCCATTAG-3 '
(SEQ ID NO: 19)
(SEQ ID NO: 2)
5'-CATTCCCAGCACCTCCTATT-3 '
(SEQ ID NO: 3)
The PCR reaction mixture contained 1.5 μL of genomic DNA (25 ng), 0.2 pmole forward and reverse primers, 200 nM dNTP, 2.5 unit Taq polymerase and 10 mM Tris-HCl (pH 9.0), 50 mM KCl , the total volume with 1.4 mM of MgCl 2 and 1X reaction buffer, 1% Triton X-100 was configured to be a 20 μL. The PCR cycling conditions of the mixture were as follows: denaturation at 94 DEG C for 5 minutes; 35 cycles of 30 seconds at a different annealing temperature for each primer set (Table 1) and 40 seconds at a primer extension of 72 DEG C; PCR Amplification was performed. The final extension was carried out at 72 ° C for 5 minutes. The electrophoresis of the amplification product was stained with ethidium bromide and performed on a 1.5% agarose gel.
Berkshire variety-specific polymorphism detection
Pig MYC sequences were retrieved from the NCBI database (Session Number: X97040.1 (Genbank)). The MYC gene is located on chromosome 4, containing three exons and a coding region of 1,359 bp, and 2,464 base pairs with 5 'UTR (untranslated region) and 3' UTR at the length of 591 bp and 514 bp, respectively . Full-length sequencing of the MYC gene was performed using 8 primer sets (P1-P8: Table 1) for DNA collected from each of the four pig breeds (Berkshire, Duroc, Landrace, Yorkshire) .
Comparing the MYC SNPs deposited in dbSNP (http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?geneId=448810), the sequencing results show that the intron 2 of the porcine MYC gene g.3350G > C polymorphism was not found in other varieties (Duroc, Landrace and Yorkshire, Table 2), but only in Berkshire varieties.
Sequencing and genotyping,
Twenty DNA samples from each of four varieties (Berkshire, Duroc, Landrace, Yorkshire) were collected and analyzed using the BigDye ® Terminator v3.1 cycle sequencing kit (Applied Biosystems, USA) and the ABI 3130 gene analyzer (Applied Biosystems, USA) Respectively. The genotyped sequence was picked and analyzed by alignment using the SeqMan 7.0 program (DNASTAR, USA) to detect the polymorphism of the gene and determine the breed specificity.
To determine the genotype from the Berkshire variety-specific polymorphism detected above (Table 2), PCR-RFLP analysis was performed on genomic DNA obtained from muscle samples amplified by PCR. The amplified PCR product (2 μL) was treated with 1 unit of Hha I restriction enzyme, 1 μL of 10 × restriction enzyme buffer and sterile distilled water to give a total volume of 10 μL. After the degradation reaction was carried out at a restriction enzyme-specific temperature and time, the DNA fragment was isolated on a 1.5% agarose gel of ethidium bromide buffer for the genotype.
The nucleotide sequence of rs321898326 is shown in SEQ ID NO: 20 in the present invention.
In the Berkshire variety MYC Genotype of g.3350G> C SNP
After polymorphic sites (g.3350G> C) were detected in the MYC gene and identified as Berkshire variety-specific, genotype analysis was performed using genomic DNA of muscle samples from the Berkshire variety. Genotypes were measured by PCR-RFLP using Hha I restriction enzyme (Table 2). The degradation of the PCR products using the Hha I restriction enzyme produced two fragments with 480 bp and 201 bp for allelotype G (G), while a fragment with 681 bp for allelotype C (Figure 1). Allelic frequencies of the GG, GC and CC genotypes of 378 breeds of Berkshire pigs were 0.399, 0.508 and 0.093, respectively (Table 3).
Polymorphic site
genotype
Number of Objects
gene
(P-value)
Genotype frequency Ob : observed genotype frequency, genotype frequency Ex : expected genotype frequency, HWE: Hardy-Weinberg Equilibrium.
Meat quality measurement
After slaughtering, the loin portion of the 10th rib of the carcass was taken and the meat quality was evaluated based on the four meat quality qualities (acidity / color, drip loss, cooking loss and shear force).
(1) pH / color
Changes in pH and color of carcass were measured at 45 and 24 hours after slaughter using a pH meter (Model 720, Thermo Orion, USA) and a colorimeter (CR-300, Minolta Camera Co, Japan). CIE L * values were read in the measurements.
(2) Drip loss
Size Standardized slices were cut from pork at 24 hours after slaughter and weighed. After weighing, the sample was suspended in an expanded polyethylene bag and stored at 2 ° C for 48 hours and then weighed again. The juice loss was calculated by the weight change rate (%).
(3) Cooking loss
After 24 hours after slaughter, a portion of the size standardized burlap sample was cut from the carcass, the sample was sealed in a hanging polyethylene bag and immersed in a heated hot pot until the internal temperature reached 72 ° C. Heat loss was measured and expressed as a percentage of the initial sample weight.
(4) Shear force
A cylindrical shearing device (1.27 cm in diameter) was used to collect the sample (parallel to the muscle fiber orientation) while heating the porcupine slurry to 72 ° C. After the sample was collected, a measuring tool (SeriesIX, Instron Crop, USA) was used to measure the maximum force (N) required to cut the cylindrical sample (perpendicular to the long axis).
Statistical analysis
The SAS 9.1 (SAS, USA) program was used to assess the genotype impact of the MYC gene on meat quality characteristics by the General Linear Model Procedure (GLM). The least significant difference (LSD) test was implemented to determine the significance between the mean values. The model was used to assess genotype effects on traits as follows:
Y ijkl = μ + Sex i + Age j + S k + G kl + e ijklm
From here,
Y ijkl = observation of each characteristic
μ = total average of each characteristic
Sex i = effect of i-th sex (i = male, female)
Age j = Age effect at the j th slaughter
S k = k-th sire effect
G l = Effect of the first genotype
e ijklm = random error
MYC Relationship between g.3350G> C SNP and meat quality characteristics
Table 4 shows the meat quality measurements obtained from 378 breeders of the breed variety in this example. The muscle samples used in this study were obtained from 138 sows and 240 birds, but no significant differences between genders were observed over meat quality qualities. Muscle pH, which affects freshness of meat due to post - metabolism, was 6.07 ± 0.30 at 45 minutes after slaughter and 5.79 ± 0.22 at 24 hours after slaughter. The color of the meat evaluated for lightness (L * ), red (a * ) and yellow (b * ) was based on CIE Lab. The lightness (L * ) was 50.49 ± 3.00, the red (a * ) was 16.36 ± 1.16, and the yellow (b * ) was 5.28 ± 1.19. Regarding the water holding capacity of meat, the juice loss was 2.51 ± 2.20 and the heating loss was 16.79 ± 3.99. The shear force was 2.66 ± 0.69. Based on the results of meat quality characteristics, g.3350G> C SNP effect was significant for pH 45 min and heat loss (Table 5, P <0.05). The Berkshire variety pig with homozygous genotype C had the highest pH of 45 min and the lowest heat loss (Table 5). Except for the two characteristics, g.3350G> C SNP did not affect pH 24h , other characteristics including meat color, juice loss and shear force (Table 5, P <0.05). There are several physico-chemical factors known to affect the quality of pork (Stalder et al., 1998). Previous studies have reported that post-hoc pH and meat quality characteristics in muscles are related (Huff-Lonergan et al., 2002). According to Jung et al. (Jung, et al., 2011), both pH 45 min and pH 24h are positively associated, while both pH 45 min and pH 24h are negative for juice loss and shear and negative for Berkshire variety pigs, It was related.
± 0.30
± 0.29
± 0.30
± 0.19
± 0.25
± 0.22
± 2.81
± 3.16
± 3.01
± 1.15
± 1.16
± 1.16
± 1.18
± 1.20
± 1.19
± 2.24
± 2.16
± 2.20
± 3.89
± 4.03
± 3.99
± 0.80
± 0.55
± 0.69
The pig MYC gene was specifically mapped to SSC4 where there is a major QTL region in the tibial region (Liu et al., 2007; Edwards et al., 2008) and daily body weight gain (de Koning et al., 2001) (Reiner et al., 1998; Cepica et al., 2003). The pig chromosome SSC 4p13 contains a synteny group with F13B, ATP1B1, GBA, ATP1A1, IVL and two microsatellites S0001 and S0067 (Andersson et al., 1994; Marklund et al , 1993).
Considering the fact that MYCs play an important role in cellular processes as a molecular hub to receive and integrate multiple signals, it is possible to control different sets of genes negatively or positively. Thus, MYC has a number of different types of cellular functions as well as one function (Potter and Marcu, 1997). In this regard, MYC expression, which is regulated by external cellular stimuli including hormones, growth factors, cytokines, lymphokines, nutritional status, development and differentiation, is associated with a variety of physiological processes (Levens et al., 1997) . More specifically, MYC has been implicated in the pathogenesis of muscle development (Miner and Wold 1991; Whitelaw and Hesketh 1992), muscle proliferation (Kipshidze et al., 2002) and lipogenesis (Ninomiya-Tsuji et al., 1993; MacDougall and Lane 1995) And thus the proto-oncogene can be a candidate gene that contributes to the diversity of obesity in pigs. In addition, previous studies have suggested the effect of MYC gene expression on glucose metabolism (Collier et al., 2003), whereby lactic acid production from the sugar chain reaction may affect the pH of the meat.
Significant differences were also found between the GG, CG and CC genotypes at pH 45 min of 5.96 ± 0.04, 5.97 ± 0.04 and 6.13 ± 0.07, respectively. Muscle pH is an important economic factor associated with the ability of muscles to retain moisture, eventually affecting the appearance of meat (Barge et al., 1991). According to Jung et al. (Jung, et al., 2011), both pH 45 min and pH 24h are positively associated, while both pH 45 min and pH 24h are negative for juice loss and shear and negative for Berkshire variety pigs, It was related. Previous studies have suggested the effect of MYC gene expression on glucose metabolism (Collier et al., 2003), whereby the lactic acid production from the sugar chain reaction may affect the pH of the meat. In this study, it was suggested that the genotype could play an influential role in the improvement of pork quality, since pH 45min was significantly higher in animals with CC genotype of g.3350G> C polymorphism than animals with other genotypes.
The structure of the muscle may or may not be affected by the heat technique, the composition of the meat and the degree of heating of the meat. Heating of meat shrinks muscle fibers, which can cause water retention and reduced heating loss, regardless of the method of heating (CHO et al., 2008). Higher heat loss is associated with lower meat quality (Schmidt et al., 2010), and higher marbling scores indicate lower heating losses (Kim and Lee, 2003). Thus, low heat loss is considered an advantageous characteristic of meat quality. Significant differences in the genetic effects of GG, CG, and CC genotypes were confirmed by measurements of 16.38 ± 0.63, 15.48 ± 0.84, and 15.50 ± 0.67, respectively, on heat loss (Table 5).
characteristic
a, b : The mean value of different characters in the same row is significantly different (P <0.05).
<110> INDUSTRIAL COOPERATION FOUNDATION CHONBUK NATIONAL UNIVERSITY <120> Method for identifying meat quality trait in berkshire using myc gene, SNP marker associated and its primer set <130> 1042522 <160> 20 <170> KoPatentin 3.0 <210> 1 <211> 6400 <212> DNA <213> Sus scrofa <400> 1 cgccacgggg cgcaaaaagg attcgtctat actgaaacct gggtgcagaa ctgggacctc 60 gggggagg cagagagtgg gtagggagcg gagagtcccc cgcagcgcgc acccgcaagc gcgctcctcc 180 tttactccag ctccggaacc ggcagaaaga tctcagggct tagcgcgtcc aggggggcaa 240 gcgggctccc gcggaggtct gtgtcggtct gcgccagcgg agcccggagg tggaggggtc 300 gaccgaggct ttggcagcga tttaggggtt tcattctaga tggaaggtgc tcagtcgagg 360 tagctgtaca cacagaatgc ctaacacccc caacaaatgc agtgggtgtt tattcagaac 420 tcgctctcca agtatacgtg gcaatgcgtt gctgagtttt ttaaaataat tccaggtacc 480 attttcctcc ctacacctct gatcattttt ccccatctac acttacccca caccacacac 540 acacaacgcg cacccgcaaa tacccccttc tttcctcctc tcttcgtagg ggttattttc 600 caaacgctgc cctttcccca gtcttaggga gggggcccct cggcccggga cgtgcgtggc 660 ttggcggtgg gtacaccgtg tattctcagt gtggagggtc agctgttccg cccgcgatga 720 tttaagcaca agggaccagt atgcggtttg tcacccagag cgctgcgcca gaggagcaag 780 cagcagagaa agagggtttg agcgggagca aaagaaaatg gtaggcgcgc gcagttaatt 840 catgcggcgc tcttactctg tttacatccg aaagctctgc gcactggagt gccaggctca 900 aagggctgag tctcctccct cccaaccacc caacccctcc ctccctcccc gggttcccaa 960 agccgagggc ggggggaaaa agaaaaaaaa gatccgctct cgcgaatccc cgcccaccgg 1020 ccctttatag gcgagggtct gcgcggctgg ggaccctccg gctgcgcatc tctcggctgc 1080 cgccgccttt gccgcacccc ggccaccgct aggctcccca ctgcctctgg aagggcaggg 1140 ctatacagag gcttggcggg aaaaagagca cggaggggag gggtgttcct agcagtataa 1200 aagccggttt tctgggcttt ctctgactcg ctgtagtaat tccagcgaga ggcagaggga 1260 gcgagcgggc gggccctcca gggtggaaga gcagagccgg ccgagcaatc tgagtcgcgc 1320 tctgggcgcc cgggggaagg gagatccgga gtgaaagagg gtcttcgcct ccgtcccggc 1380 cgcccccacc ccaccctgcc cgccgacccc tgccagcggt ccgccacccg cgccgcatcc 1440 acgaaacttt gcccactgca gcgggcgggt actttccact ggaacttaca acacccgagc 1500 gacaacgcga ctctccggac gcggagaggc tattctgcct atttggggag acacttttcc 1560 ctgtcgctgc ccacgactcg ctcctctgaa aggcgctcct cgccgctttt tggacgctgg 1620 atttccttcg gatagtggaa aacccggtga gcacccagac ctatttgtct tttaatttct 1680 tcttgctacc gctttcaatc cgcgatgagt cgaatgcccg aattggggtg tcttttctcc 1740 cattcctgct ctattgacac ttttctcaga gtagttgtgg ttggtcgggg tagggtgggg 1800 acgaaccaga gctgggtcgg ggtgcaatga cttgtcaaga aggaagagag gaaggcagag 1860 ggagaacggg gtgctttttt ttgaaagtag cctttagagg ggttggttgt ttttgtgtat 1920 ttttgtttta atgtagatga acgctgacct cggccggctg ggcattcttg ctttattgca 1980 tttgattgct tttttgcgt ttggagggag aaggggggta tgcttcgcag tgggcagaaa 2040 accctttacg tcctgagctc ctatgagtaa gaattacata ttgcgtgtgg agcgagctcc 2100 gcagcctctg acttttttgc cgtctccgaa agggcattta aatttcggct taccgcattt 2160 gtgacagcgg agactgcggc gagtcccgcc cgccgatccc cgcggcgctt caattcgccc 2220 cccggctcct cttagaagtt ggcgtttggc tttttaaaag aataatgaaa ttaaagctct 2280 tggtcctcgg agatgttaag acgtggtgtt ggggaagcgc ggggacgggg gaaaaggagg 2340 gtggcgcaaa atgtgtccga ttctcctgga attattgact cgggggaaaa cgaggacaaa 2400 tctccgcacc ccgccttggc tcgtcggccg cctctggtgt ccccgctcgg gtgtccctgc 2460 gatgcggagg gactgcgggc agcggggttc ggggcttttc cagaacagct gctaccctcg 2520 gcggcgggag aggggaagac gcccggcttc gggcgcggaa ttgcagcagg gtctctggcg 2580 cagttgcgtc gccgtactgg gtgttgcaag gaggtgtctg ttatttaaca gaccccccac 2640 acaccctttc attggtcgga ggtgggggtt aaggcccgaa gactgagctc tctaccccag 2700 ccggagagag aaaagaagaa aagccggcaa aggaaggaag gggggcgcgc ggggggtggg 2760 gatggaggag gcggagaggg gaggttggga ggggcggtgg cgccggcggg ggaaggagcg 2820 cggcgagggc gcgagtggga acgggcgctg cggaggggcc ctgtcgcggg aggggggccg 2880 cgcgcatccg ctcccgccgc ggcgcctctt cgccttcccc ttcaggtggc gcaaaacttt 2940 gcgccttggc gtttggcaga ttgtatttcc ccaccgccgc ctttccctgg cctgttaaag 3000 gaggctaaag ctgatttcga atgccaggcg gccgcggggc agactccccg gtttcgcgtt 3060 ccgggctccc gggggaacgg aagctccgtg ggcgccacag aggtggttca ccaagtgtgt 3120 gtctgagata gtagagggtc acttcgaggg ggtggaaggg tgcctttttg ttaatccccc 3180 actcccaccg gccgctttgg gggatagcgc tggaagggga gtggttcccg atcgtggcgc 3240 gcagtgagcg cggcgcgggg tttccacccg aaaaaaagac cccattaact caagacgcgc 3300 ccccctcttt gtgtgtcccc tccagcaggc tgccgcgatg cccctcaacg tcagcttcac 3360 caacaggaac tatgacctcg actacgactc ggtgcagcct tatttctact gcgacgagga 3420 ggagaacttc taccagcagc agcagcagag cgagctgcag ccgccggcac ccagcgagga 3480 tatcggaag aaattcgagt tgctgcccac cccgcccctg tccccgagcc gccgctccgg 3540 gctctgctcg ccctcgtacg tcgcggtcgc gtccttctcc cccaggggag acgatgacgg 3600 cggcggcggc agcttttcca cggccgacca gttggagatg gtgaccgagc tgctgggagg 3660 agacatggtg aaccagagct tcatctgcga ccccgacgac gagaccttca tcaaaaacat 3720 tattatccag gactgtatgt ggagcggctt ctcagccgcc gccaagctcg tgtcagagaa 3780 gctggcctcc taccaggctg cgcgcaaaga cagcggcagc ccgattcccg cccgcgggca 3840 cggcggctac tccacctcca gcttgtacct gcaggacctg agcgccgccg cctccgaatg 3900 catcgacccc tcggtggtct tcccctaccc gctcaacgac agcagctcgc ccaagccctg 3960 cgcctccccc gactccaccg ccttctcccc gtcctcggac tctctgctct cctccgccga 4020 gtcctcccca cggggcagtc ccgaaccctt ggctctccac gaggagacac cacccaccac 4080 cagcagcgac tctggtaagc tgggcccctc caagcccgtc agaggggttg gctggctgtc 4140 tttcctagtc tcatgggcct cggattcaaa gggccatttc agttgtccct gccttttctc 4200 ttctcttcca tttggaaaag aaatgcagat cctgatggag gaactcatca ttcttgaacg 4260 ctgggcttca gcgttccacc catcccttcc acccatccct tccccggaga cttccctagt 4320 gccagcctcc tcctttccct ccctttgaga atttcattgg ggttttaaaa tctttatcta 4380 gccactccct ccgctccctc ttacccttct taagcatttt aattaccctg gggggtgggg 4440 aggttagctc tcctggttaa gagacctatc tcttagagtg aatgaattgc ctcccttaac 4500 ttcttagaag ttggtggcct ttagtggacg acagatcaac aaaaataaag gggggtggtg 4560 gacagagagc aggcagcctc ccacctgggt accaagagct agtgaaagtg ccttaaaagt 4620 ggatgggctg aggagctggg atcttttcag cctcttttga acacttaaaa gcaagtcctt 4680 tccaaaattg ggcttctttt ctccctccct caccccctag gacttttggc aaagctgcaa 4740 gacttttttt tttttttttg cccttccagt aaaataggga gttggtaaag tcataccaaa 4800 gatttgcagc tattgtttgc aacacctgaa gggttcttgg taaagtccct tgcaaatagg 4860 aggtgctggg aatgtgcttt gctttgggtg tgttcagagc ctcattaagt cttaggtaag 4920 aattggcatc aatgttgtaa atggtaaatt gtaattttct tgtctgtgcc ataaacccag 4980 ctgtcatttt cctccctgag actctgccat catgggaaga agggcacagg tgattggcta 5040 gttctctggg gataactctg tgccagggtc ctttcttacc taatgctgcc ttatatttgc 5100 atggtattaa tttaatctgg tccttgattg ctttaaggaa accagttgct aattgggtga 5160 tttctctttc ctttcttaaa gaggaggaac aagaagatga ggaagaaatt gatgttgttt 5220 ctgtggaaaa gaggcagccc cctgccaaaa ggtcggaatc ggggtcaccc tctgctggag 5280 gccacagcaa acctcctcac agccccctgg tccttaagag atgccatgtg tccacgcacc 5340 agcacaatta tgcagcgccc ccctccacca ggaaggacta tccctctgcc aagagggcta 5400 agttggacag tggcagggtc ctgaaacaga tcagcaacaa ccgaaaatgc gccagcccca 5460 ggtcctcgga cacggaggag aatgacaaga ggcggacaca caatgtcttg gaacgccaga 5520 ggagaaacga gctgaaacgc agcttttttg cccgtcgaga ccagatccca gagttggaga 5580 acaatgaaaa agcccccaag gtagttatcc ttaaaaaagc cacagcttac atcctgtctg 5640 tccaagcaga ggagcaaaag ctcgtgtcag aaaaagacgt gctgcggaag aggcgagaac 5700 agttgaaact caaacttgaa cagctacgga actcttgccc ataaattgac ctattggagg 5760 gaggaactag aatggcccat gaattcacac ttgttactaa gagaaagtaa ggaagaaggt 5820 tcctttggac agaactgctt gctggccatt atttgaactc attattacgt atgaactttt 5880 tctcgaatgc atgatccagt gcaacctcac aaccttggct gggtcttggg actaaaaggc 5940 ctagacataa tataaactgc ctcaaatgga actttgagca taaaagaact ttttttatgc 6000 ttaccatctt tttttttttc ctttaacaga tttgtattta aaaattgttt ttacaaaatc 6060 ttaatgttca gcaaactttc ctctgtaaat atggtcatta aatgtacata attttaataa 6120 aaaacgttta tagcaattat acaaaatttc aacaagacat gtattataaa ccataatttt 6180 ttttatttaa gtaccttttc ctttttaaag ttgatttttt tctattgttt ttagaaaaaa 6240 ataaaatatt tggcaaatat ataattaagc caaatgttaa gttgtgagtg ttttgtttct 6300 tttctttttt ttttttcttc ctctttctct tttcatcaat tacaaagtaa cggaatttgg 6360 cccttaatgg agttgagcct acaaagatgg gaaaagaagt 6400 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> RP1 forward primer <400> 2 cagttgtccc tgccttttct 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> RP1 reverse primer <400> 3 cattcccagc acctcctatt 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P1 forward primer <400> 4 ggttttctgg gctttctctg 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P1 reverse primer <400> 5 caaaaacaac caacccctct 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P2 forward primer <400> 6 tcttcttgct accgctttca 20 <210> 7 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> P2 reverse primer <400> 7 gggtagcagc tgttctgga 19 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P3 forward primer <400> 8 ttattgactc gggggaaaac 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P3 reverse primer <400> 9 cacacttggt gaaccacctc 20 <210> 10 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> P4 forward primer <400> 10 ggcgtttggc agattgtat 19 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P4 reverse primer <400> 11 gtaggaggcc agcttctctg 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P5 forward primer <400> 12 tgaaccagag cttcatctgc 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P5 reverse primer <400> 13 aagggaggca attcattcac 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P6 forward primer <400> 14 cgctccctct tacccttctt 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P6 reverse primer <400> 15 accctggcac agagttatcc 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P7 forward primer <400> 16 gaagaagggc acaggtgatt 20 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P7 reverse primer <400> 17 actggatcat gcattcgaga 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P8 forward primer <400> 18 cggaactctt gcccataaat 20 <210> 19 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> P8 reverse primer <400> 19 ttaaggccca gacccattag 20 <210> 20 <211> 51 <212> DNA <213> Sus scrofa <220> <221> misc_feature (26) <223> n = g or c <400> 20 tgcttaagaa gggtaagagg gagcgnaggg agtggctaga taaagatttt a 51
Claims (8)
(a) amplifying a MYC gene comprising the SNP marker of claim 2 from a genomic DNA of a sample isolated from a Berkshire variety pig;
(b) amplifying the MYC gene amplified in step (a) with restriction enzyme Hha I ; And
(c) comparing the polymorphism (RFLP) in the gene fragments obtained in the step (b).
(a) amplifying a MYC gene comprising the SNP marker of claim 2 from a genomic DNA of a sample isolated from a Berkshire variety pig;
(b) amplifying the MYC gene amplified in step (a) with restriction enzyme Hha I ; And
(c) comparing the polymorphism (RFLP) in the gene fragments obtained in the step (b).
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