KR101273333B1 - A new primer set for identifying fatty acid composition in broiler chicken and a method of selecting chicken by using the said primer set - Google Patents
A new primer set for identifying fatty acid composition in broiler chicken and a method of selecting chicken by using the said primer set Download PDFInfo
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Abstract
The present invention is a homozygote of a single base polymorph of the exon No. 2 of the chicken selection primer set having a high unsaturated fatty acid content consisting of the nucleotide sequence of SEQ ID NO: 1 and the nucleotide sequence of SEQ ID NO: 2, SCD (Stearoyl-CoA desaturase) gene of chicken A method of selecting a chicken having a high content of unsaturated fatty acids, including determining whether the genotype is a genotype or a primer set consisting of a nucleotide sequence of SEQ ID NO: 1 and a base sequence of SEQ ID NO: 2, and detecting a gene marker of a chicken having a high content of unsaturated fatty acids It relates to a chicken discrimination kit having a high unsaturated fatty acid content comprising a detection means.
Description
The present invention relates to a genetic marker for identifying fatty acid composition of chicken, specifically broiler chicken, that is, a novel primer set and a method for screening broiler chicken meat using the same.
Fatty acids ingested by humans are known to cause high cholesterol and increase blood glucose levels, and thus affect cardiovascular and metabolic diseases. In particular, excessive intake of saturated fatty acids (SFA), such as lauric acid (C12: 0), myritic acid (C14: 0), or palmitic acid (C16: 0), is closely related to cardiovascular disease. It is reported. On the other hand, high intakes of polyunsaturated fatty acid (PUFA) or monounsaturated fatty acid (MUFA) increase the activity of low-density lipoprotein (LDL) receptors and reduce the circulating concentration of LDL cholesterol. Is being reported.
In recent years, the emphasis on health-oriented lifestyles, for consumers with a high health concern, the composition of fatty acids in the selection of meat or meat plays a very important factor, and according to these needs to find meat with a good fatty acid composition Research continues to be important.
In this respect, poultry meat is healthy in that it contains a lower percentage of saturated fatty acids and higher polyunsaturated fatty acids, especially long-chain omega-3 eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). It is known to be oriented. In particular, red meat of chicken has been reported to reduce apolipoprotein B and total cholesterol levels in the body as a dietary replacement, and health-oriented consumers are replacing white meat with red meat. .
Therefore, the production and selection of livestock, especially poultry, which has been of high quality meat, including the preferred fatty acid composition, has become an important objective for producers and researchers.
As a method for selection and production of animals having a desirable fatty acid composition, marker-assisted selection (MAS) has been provided by using a marker or primer set capable of investigating genotype. Specifically, the marker help selection method has been applied in the breeding industry by directly selecting a gene that affects the fatty acid composition.
For example, studies are being conducted to select individuals having a high specific fatty acid composition by using a marker or primer set capable of identifying a genotype of a specific gene that affects a fatty acid composition such as SCD (Stearoyl-CoA desaturase).
The SCD is an enzyme that promotes the conversion of saturated fatty acid to monovalent saturated fatty acid (MUFA) in mammalian adipocytes, and can promote delta (Δ) 9 unsaturated reaction of saturated fatty acid and monovalent saturated fatty acid, and the delta of fatty acid substrate. The insertion of cis-double bonds at position 9 is known to contribute to the synthesis of unsaturated fatty acids.
However, these SCD genes are mainly studied in relation to cows, and it has been reported that the fatty acid ancestors of cows are closely related to the SCD genotype. In addition, recent studies in pigs have reported that the SCD gene has a significant effect on fatty acid composition and melting point.
However, there have been no reports on the effect of SCD gene on fatty acid composition in chicken, and there is no study on the association between specific genotype or polymorphism of SCD and fatty acid composition in chicken. .
The present invention, as described above, in order to solve the problems of existing broiler production farms by selecting broilers that can produce a high quality chicken meat with a high unsaturated fatty acid content, gene for confirming the fatty acid composition of chicken from many broilers It is an object of the present invention to provide a marker, more specifically, a gene marker for quickly and accurately identifying chickens having a high content of unsaturated fatty acids.
In addition, the present invention aims to provide a method for selecting chickens having a high content of unsaturated fatty acids using the above gene markers.
In order to achieve the above object, the present invention provides a screening primer set capable of selecting a chicken having a high unsaturated fatty acid content.
In addition, the present invention provides a selection method that can select a chicken having a high unsaturated fatty acid content. The screening method may be to use a screening primer set that can screen the chicken with a high content of unsaturated fatty acids.
In addition, the present invention provides a selection kit that can select a chicken having a high unsaturated fatty acid content. The selection kit may include a selection primer set for selecting chickens having a high content of unsaturated fatty acids.
The present inventors conducted research to find genetic markers related to fatty acid composition of chickens, specifically, genetic markers capable of selecting chickens having high unsaturated fatty acid content, and thus, five single-base polymorphisms in chicken SCD (Stearoyl-CoA desaturase) genes. SingleNucleotide Polymorphism (SNP) located in Exon 2 is present in Palmitoleic acid (C16: 1), Palmitic acid (C16: 0) and Myristoleic acid (Myristoleic). acid, C14: 1) was confirmed to have a high correlation with the homozygous genotype using a novel primer that can identify the single base polymorphism located in the exon 2 from this, quickly and accurately unsaturated The present invention was completed by confirming that chickens capable of producing chicken having a high fatty acid content can be selected.
Hereinafter, the present invention will be described in detail.
In one aspect of the invention, the present invention relates to a primer set for chicken selection having a high unsaturated fatty acid content.
The primer set has a high correlation with the unsaturated fatty acid content, in particular with respect to the content of palmitoleic acid (C16: 1), palmitic acid (C16: 0) and myristoleic acid (C14: 1). There are markers specifically, it is possible to select chickens with a high unsaturated fatty acid content using a genetic marker. The genetic marker may be related to the single nucleotide polymorphism of exon 2 of the chicken SCD (Stearoyl-CoA desaturase) gene.
In this aspect, the primer set may be composed of the nucleotide sequence of SEQ ID NO: 1 and the nucleotide sequence of SEQ ID NO: 2.
Specifically, the chicken having a high unsaturated fatty acid content may be a broiler chicken having a monobasic polymorph of exon 2 of the SCD (Stearoyl-CoA desaturase) gene having a homozygous genotype.
In another aspect of the present invention, the present invention relates to a method for selecting chickens having a high unsaturated fatty acid content, specifically a method for selecting a broiler chicken capable of producing a chicken having a high unsaturated fatty acid content.
The selection method is a marker-assisted selection (MAS) using a marker or primer set capable of investigating the genotype, specifically, a marker or primer set related to a chicken SCD (Stearoyl-CoA desaturase) gene. Marker help selection.
In this aspect, the method of selecting a chicken having a high content of unsaturated fatty acids may include determining whether the single nucleotide polymorphism of exon 2 of the SCD (Stearoyl-CoA desaturase) gene of the chicken is homozygous genotype. .
Specifically, the step of determining whether the homozygous genotype may be characterized by using a primer set consisting of the nucleotide sequence of SEQ ID NO: 1 and the nucleotide sequence of SEQ ID NO: 2. The homozygous genotype of the single base polymorph of exon 2 of the SCD gene of the chicken may be AA or GG type. As an example, the homozygous genotype of the single base polymorph may be GG type.
In addition, the present invention comprises the steps of a) collecting a DNA sample from the chicken to be identified; b) amplifying the DNA sample using the DNA sample and a primer set consisting of SEQ ID NO: 1 and SEQ ID NO: 2; c) treating the amplification product with restriction enzymes; And d) may be a method of selecting a chicken having a high unsaturated fatty acid content comprising the step of confirming the genotype by electrophoresis of the treatment treated with the restriction enzyme.
The DNA sample may be genomic DNA of chickens taken from the chicken, specifically broiler chickens to be searched. More specifically, SCD (Stearoyl-CoA desaturase) gene of chicken isolated from the genomic DNA. It may be a site corresponding to exon No. 2 or exon No. 2 of the SCD (Stearoyl-CoA desaturase) gene of the chicken isolated from the genomic DNA.
The DNA sample may be a DNA sample taken from a chicken, specifically broiler chicken, more specifically, genomic DNA (genomic DNA) obtained from chicken blood, semen, gravy or chicken tissue, preferably chicken Genomic DNA from tissues of
Determining whether the single base polymorphism of exon 2 of the chicken SCD (Stearoyl-CoA desaturase) gene of the chicken is a homozygous genotype is a DNA sample and a primer set consisting of SEQ ID NO: 1 and SEQ ID NO: 2 Amplifying the DNA sample using; Treating the amplification product with restriction enzymes; And it can be carried out by a method comprising the step of confirming the genotype by electrophoresis of the treatment treated with the restriction enzyme.
In addition, the step of confirming whether the single base polymorphism of the exon 2 of the chicken SCD (Stearoyl-CoA desaturase) gene is homozygous genotype is a marker help selection method using a marker or primer set that can examine the genotype It can be done with
In the marker help selection method, the DNA sample of the chicken is used as a template, the primer set consisting of SEQ ID NO: 1 and SEQ ID NO: 2 is performed as a primer, and the sequencing of the PCR reaction product is performed. After checking or treating the PCR reaction product with a restriction enzyme, electrophoresis using an agarose gel and a polyacrylamide gel, a DNA band resulting from the electrophoresis (DNA band) This can be done by checking).
The amplifying may be performed by PCR using a DNA set as a template and a primer set consisting of the nucleotide sequence of SEQ ID NO: 1 and the nucleotide sequence of SEQ ID NO: 2 as a primer. Specifically, the amplifying step is a PCR using a primer set consisting of the nucleotide sequence of SEQ ID NO: 1 and the nucleotide sequence of SEQ ID NO: 2 as a template using exon 2 of the chicken SCD (Stearoyl-CoA desaturase) gene as a template. This can be done via reaction.
The restriction enzyme used in the restriction enzyme treatment of the amplification product may be Aci I.
Specifically, the genotype may be a monobasic polymorph of exon 2 of the SCD (Stearoyl-CoA desaturase) gene of chicken. In addition, the step of identifying the genotype may be performed by a method of confirming that the single nucleotide polymorphism of exon 2 of the SCD (Stearoyl-CoA desaturase) gene of the chicken is homozygous genotype. The homozygous genotype may be type AA or type GG, for example, type GG.
In an embodiment of the present invention, electrophoresis of the PCR reaction product of a sample taken from a chicken with a restriction enzyme showed that the single nucleotide polymorphism of No. 2 exon of chicken SCD (Stearoyl-CoA desaturase) gene was homozygous. In the case of the conjugate genotype, two bands of 300 bp to 350 bp size, specifically, 325 bp to 330 bp size and 120 bp to 160 bp size, specifically 135 bp to 145 bp size are identified, or 220 bands are identified. bp to 250 bp size, specifically 230 bp to 240 bp size band, 120 bp to 160 bp size band, specifically 135 bp to 145 bp size band and 80 bp to 100 bp size band, specifically 90 bp to 95 bp size band 3 can be confirmed.
For example, if you want to select a broiler that produces a chicken meat containing a large amount of palmitoleic acid, three bands of 220 bp to 250 bp, 120 bp to 160 bp and 80 bp to 100 bp are identified. You can choose chicken.
In another aspect of the present invention, the present invention is an unsaturated fatty acid comprising a primer set consisting of a nucleotide sequence of SEQ ID NO: 1 and a nucleotide sequence of SEQ ID NO: 2 and a detection means for detecting a gene marker of a chicken having a high unsaturated fatty acid content. It may be a high content chicken discrimination kit.
The chicken discrimination kit may further include a DNA sample collected from the chicken to be discriminated.
Gene expression of chickens having a high content of unsaturated fatty acids may be a homozygous genotype of a single base polymorph of No. 2 exon of the SCD gene of the chicken, the homozygous genotype may be AA or GG type, for example, GG type.
The detection means may be a means for detecting a conventional gene marker, preferably may include a means for detecting the gene marker by performing a PCR reaction, restriction enzyme treatment reaction and electrophoresis.
Specifically, the detection means may be a means for performing a PCR reaction using the DNA sample and primer set; Means for treating the product of the PCR reaction with a restriction enzyme; And it may be a detection means comprising a means for confirming the genotype by electrophoresis the treatment treated with the restriction enzyme. The restriction enzyme may be Aci I.
The means for identifying the genotype by electrophoresis may be a means for confirming the number and size of DNA bands resulting from the electrophoresis after electrophoresis using an agarose gel or polyacrylamide gel.
In an embodiment of the present invention, electrophoresis of the PCR reaction product of a sample taken from a chicken with a restriction enzyme showed that the single nucleotide polymorphism of No. 2 exon of chicken SCD (Stearoyl-CoA desaturase) gene was homozygous. In the case of the conjugate genotype, two bands of 300 bp to 350 bp size, specifically 325 bp to 330 bp size and 120 bp to 160 bp size, specifically 135 bp to 145 bp size are identified, or 220 bp bands are identified. 3 to 250 bp band, specifically 230 bp to 240 bp band, 120 bp to 160 bp band, specifically 135 bp to 145 bp band and 80 bp to 100 bp band, specifically 90 bp to 95 bp band Can be identified.
For example, if you want to select a broiler that produces a chicken meat containing a large amount of palmitoleic acid, three bands of 220 bp to 250 bp, 120 bp to 160 bp and 80 bp to 100 bp are identified. You can choose chicken.
The method for analyzing monobasic polymorphisms of chicken SCD gene using the primer set of the present invention serves as an indicator for the unsaturated fatty acid content of the chicken, which makes it possible to easily and quickly select chickens having an excellent fatty acid composition. As it enables the production and selection of chickens with high quality meat with composition, it helps to improve the competitiveness and efficiency of breeding farmers who need international competitiveness by trade treaties such as FTA, thereby improving the profitability of farmers raising broilers. And sales growth.
1 is a graph confirming g.3728A> G single nucleotide polymorphism of SCD gene No. 2 exon based on sequencing among genomic DNAs of chickens obtained from a broiler chicken subject according to an embodiment of the present invention. Figure 1a is the result of confirming the broiler chicken named P2R, Figure 1b is the result of confirming the broiler chicken named P2G.
Figure 2 is a sequence of sequencing (sequencing) the SCD gene No. 2 exon in genomic DNA of chickens collected from the experimental broiler chickens according to an embodiment of the present invention. The left side of the base sequence means a sample, that is, the type of broiler broiler collected, and the number on the right side indicates the position (bp).
3 is a photograph showing a genotyping result showing amplification of the SCD gene No. 2 exon of broiler chickens using a primer of the present invention and enzyme treatment (PCR-RFLP) according to an embodiment of the present invention. The numbers on the left represent the size (bp), M represents the marker, and AA, GA and GG each represent the polymorphism.
Hereinafter, the present invention will be described in more detail with reference to production examples and examples. However, the following Preparation Examples and Examples are merely illustrative examples for the purpose of facilitating understanding of the present invention, and can be modified into various other forms, and the scope of the present invention is not limited to the following Examples.
Example
95 Ross broiler chickens were reared at 28 weeks of age until the carcass weight was greater than 1.6 kg. After sacrificing the broiler chicken at 28 weeks of age, the thigh muscles were separated, and the tissue obtained from the muscle was used as a sample, and DNA extraction and fatty acid composition analysis were performed.
In addition, as a result of the above analysis, specifically, the statistical analysis of the correlation between the SCD genotype and fatty acid composition was performed using the GLM (general linear model) in the SPSS version 17.0 program (SPSS, USA).
The significance between the genotype was performed by Tukey HSD test, and specifically, the correlation between genotype and fatty acid composition was analyzed using a statistical model described in the following formula. In the following formula, Y is the phenotype data (fatty acid composition) of sample i, μ is the overall mean, G is the genotype of sample i, and ε means error.
[formula]
Y i = μ + G i + ε i
Example 1. Fatty acid analysis
The fatty acid analysis was performed in the following manner. First, the whole lipid was extracted from the sample using a mixed solution in which chloroform and methanol were mixed at a volume ratio of 2: 1 according to the experimental procedure reported by Folch et al. (1957). In addition, fatty acid methyl esters (FAME) were separated from lipids extracted with BF3-methanol (Sigma-Aldrich, USA) using HP-6890N gas chromatography.
Specifically, the lipid extracted using the BF3-methanol is split into a crack inlet having a split ratio of 50: 1 on an Omegawax 320 capillary column (30 mx 0.25 mm x 0.25 μm). inlet, Supelco, USA). The temperature of the ramp oven was adjusted as follows. First, hold at 150 ° C. for 3 minutes, increase to 2.5 ° C./min until reaching 180 ° C., then hold for 5 minutes, then increase to 2.5 ° C./min until reach 220 ° C., and then 25 minutes It was carried out by the method of maintaining. The temperature of the crack inlet was adjusted to 210 ° C. The air maintained a flow of 0.7 mL / min as carrier gas.
The average fatty acid composition of the broiler 95 heads confirmed from the analysis results is shown in Table 1 below. Table 1 shows the percentage and standard deviation of the analyzed average fatty acid composition, SFA in Table 1 means saturated fatty acids, MUFA means monounsaturated fatty acids, PUFA means polyunsaturated fatty acids.
As shown in Table 1, the fatty acid composition in broiler chicken was confirmed that the level of saturated fatty acids appeared at a lower level compared to beef. In this regard, excessive intake of saturated fatty acids can result in enhancement of plasma cholesterol, which can lead to cardiovascular disease, in particular lauric acid (C12: 0), myristic acid (C14: 0) and palmitic Since saturated fatty acids such as acid (C16: 0) are known to be the most harmful fatty acids for cardiovascular diseases, the association between the saturated fatty acids and the broiler genotype was examined.
Example 2. Analysis of association between genotype and fatty acid
In order to compare the fatty acid composition and genotype analyzed in Example 1, first, 15 of the 95 numbers were selected, DNA samples were obtained from the thigh muscles, and a single nucleotide polymorphism was confirmed.
Specifically, genomic DNA samples were subjected to phenol extraction on muscle samples, 20 mg / ml proteinase K digestion, and then Prime Prep ™. Genomic DNA was extracted by using Genomic DNA Isolation Kit (GenNetBio, Korea).
Using the extracted DNA sample as a template and using a primer pair prepared based on the nucleotide sequence of the chicken SCD gene of Table 2 (GenBank Acc. No NC_006093) as a primer, PCR reaction was performed under the conditions of Table 3 below. It was. The PCR reaction solution for the PCR reaction is 50 ng of each genomic DNA sample, 10X buffer buffer (reaction buffer, 100 mM Tris-Cl, 400 mL KCl, 20 mM MgCl2, GenetBio, South Korea) Taq DNA polymerase, 10 mM dNTP And the primers of Table 2 below. The PCR reaction was performed using a GeneAmp PCR System 2700 (Applied Biosystems, US) or C1000 ™ Thermal Cycler (Thermal Cycler, BioRad, USA). As a result of the PCR, it was confirmed that the fragment of 468 bp size was amplified.
After completing the PCR reaction, the amplification products were observed on a 1.5% standard agarose gel stained with ethidium bromide (EtBr, GenetBio, South Korea). Each PCR product was purified using AccuPrep PCR Purification Kit (Bioneer, Korea). Purified PCR products were sequenced using the same primers as the PCR reaction on an automated 3730 XL DNA sequencer (Applied Biosystems, USA). DNA sequence is BioEdit program ver. 7.00 (Tom Hall, Ibis Therapeutics, USA) and single nucleotide polymorphism (SNP) were identified based on electrophoretic polymorphisms. The results confirmed above are shown in FIGS. 1 and 2.
As a result, five monobasic polymorphisms were identified in exons (exons 2 and 4) and introns (introns 2 and 5) of the SCD gene. In particular, the single base polymorphism identified in exon 2 was selected and compared for each sample.
As shown in FIG. 1, the g.37284A> G monobasic polymorphism located at SCD gene exon 2 contains a restriction site that can be recognized by restriction enzymes, thereby facilitating genotyping. PCR-RFLP method has been confirmed that can be distinguished from individual to specific base at a specific base position, that is, A or G.
In addition, as shown in Figure 2, the PCR amplification product of the amplified 468 bp SCD gene was cleaved using Aci I restriction enzyme, the individual with
14 fatty acid compositions of the muscle samples of each of the AA, GG and GA types of chickens were analyzed, and correlation analysis between gene polymorphism and fatty acid traits was performed using the statistical analysis method.
The results are shown in Table 4 below. P-values in Table 4 below refer to the difference in phenotype mean between genetic polymorphism and fatty acid composition trait. In addition, Table 4 shows the percentage and standard deviation of each fatty acid composition analyzed, SFA in Table 4 means saturated fatty acids, MUFA means monounsaturated fatty acids, PUFA means polyunsaturated fatty acids. . Mean values with different superscripts such as a or b in the same column of Table 4 mean that they are significantly different in P <0.01 (Tukey's test).
As shown in Table 4, g.37284A> G monobasic polymorphism of the SCD gene is mystoleic acid (C14: 1), palmitic acid (C16: 0), palmitoleic acid (C16: 1) and saturated fatty acid ( SFA). Specifically, the heterozygous genotype (GA) of the monobasic polymorphic genotype was significantly increased compared to the homozygous genotype, and palmitic acid and saturated fatty acid were significantly increased, and myristoleic acid was reduced.
In addition, the effect of increasing palmitoleic acid in individuals with GG among homozygous genotypes has been confirmed. The palmitoleic acid is known as a fatty acid having a good effect of reducing harmful cholesterol, has the effect of reducing the deposition of fat in the blood vessels, reducing the aggregation of platelets. Therefore, in relation to healthy meat intake, the single base polymorph of exon 2 of the chicken SCD (Stearoyl-CoA desaturase) gene in chickens has a high palmitoleic acid content. It is expected to be preferable in terms of selection of individuals.
<110> Foundation of University-Industry Research Collaboration <120> A NEW PRIMER SET FOR IDENTIFYING FATTY ACID COMPOSITION IN BROILER CHICKEN AND A METHOD OF SELECTING CHICKEN BY USING THE SAID PRIMER SET <130> DP20120142 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> SCD-F <400> 1 cccccagaaa gaaaaagtcc 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> SCD-R <400> 2 caaaaatccc acccaacaac 20
Claims (16)
b) amplifying the DNA sample using a primer set consisting of the DNA sample and the nucleotide sequence of SEQ ID NO: 1 and the nucleotide sequence of SEQ ID NO: 2;
c) treating said amplification product with restriction enzyme Aci I; And
d) electrophoresis of the treatment treated with the restriction enzyme Aci I to determine whether the single nucleotide polymorphism of No. 2 exon of the SCD (Stearoyl-CoA desaturase) gene of the chicken is homozygous genotype;
A method of screening for chickens having a higher unsaturated fatty acid content than a chicken having a heterozygous genotype of the exon 2 polysaccharide of the SCD (Stearoyl-CoA desaturase) gene of a chicken.
The amplifying step includes the DNA sample as a template, and performing a PCR reaction using a primer set consisting of the nucleotide sequence of SEQ ID NO: 1 and the nucleotide sequence of SEQ ID NO: 2 as a primer.
A method of screening for chickens having a higher unsaturated fatty acid content than a chicken having a heterozygous genotype of the exon 2 polysaccharide of the SCD (Stearoyl-CoA desaturase) gene of a chicken.
The homozygous genotype is AA or GG type
A method of screening for chickens having a higher unsaturated fatty acid content than a chicken having a heterozygous genotype of the exon 2 polysaccharide of the SCD (Stearoyl-CoA desaturase) gene of a chicken.
The homozygous genotype is GG type
A method of screening for chickens having a higher unsaturated fatty acid content than a chicken having a heterozygous genotype of the exon 2 polysaccharide of the SCD (Stearoyl-CoA desaturase) gene of a chicken.
Amplification means for amplifying the DNA sample using a DNA sample collected from a chicken to be discriminated and a primer set consisting of the nucleotide sequence of SEQ ID NO: 1 and the nucleotide sequence of SEQ ID NO: 2;
Processing means capable of processing the DNA sample amplified by the amplification means with restriction enzyme Aci I; And
Detection means for detecting whether the single nucleotide polymorphism of exon 2 of the chicken SCD (Stearoyl-CoA desaturase) gene is homozygous genotype using the treatment treated with the restriction enzyme Aci I
Containing
A chicken discrimination kit having a monobasic polymorph of exon No. 2 of the SCD (Stearoyl-CoA desaturase) gene of a chicken having a higher unsaturated fatty acid content than a chicken having a heterozygous genotype.
The homozygous genotype is characterized in that the type AA or GG
A chicken discrimination kit having a monobasic polymorph of exon No. 2 of the SCD (Stearoyl-CoA desaturase) gene of a chicken having a higher unsaturated fatty acid content than a chicken having a heterozygous genotype.
Detection means for detecting whether the homozygous genotype is characterized in that the means for confirming the genotype by electrophoresis of the treatment treated with the restriction enzyme
A chicken discrimination kit having a monobasic polymorph of exon No. 2 of the SCD (Stearoyl-CoA desaturase) gene of a chicken having a higher unsaturated fatty acid content than a chicken having a heterozygous genotype.
The means for identifying the genotype by electrophoresis is a means for confirming the number and size of DNA bands resulting from the electrophoresis after electrophoresis using an agarose gel or polyacrylamide gel.
A chicken discrimination kit having a monobasic polymorph of exon No. 2 of the SCD (Stearoyl-CoA desaturase) gene of a chicken having a higher unsaturated fatty acid content than a chicken having a heterozygous genotype.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100822741B1 (en) | 2005-12-21 | 2008-04-17 | 대한민국 | DNA marker for identification of intramuscular fat marbling in Hanwoo and Method of identification of intramuscular fat in Hanwoo by using the same |
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|---|---|---|---|---|
| KR100822741B1 (en) | 2005-12-21 | 2008-04-17 | 대한민국 | DNA marker for identification of intramuscular fat marbling in Hanwoo and Method of identification of intramuscular fat in Hanwoo by using the same |
Non-Patent Citations (2)
| Title |
|---|
| Mol. Biol. Rep, Vol. 39, pp. 2677-2682 (2011.06.21.) * |
| Mol. Biol. Rep, Vol. 39, pp. 3153-3160 (2011.07.15.) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115807100A (en) * | 2022-07-22 | 2023-03-17 | 华南农业大学 | SNP molecular marker related to broiler abdominal fat percentage and application thereof |
| CN115807100B (en) * | 2022-07-22 | 2023-08-22 | 华南农业大学 | SNP molecular marker related to abdominal fat rate of broiler chickens and application thereof |
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