CN113789392A - SNP marker related to growth of channel catfish and application thereof - Google Patents

SNP marker related to growth of channel catfish and application thereof Download PDF

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CN113789392A
CN113789392A CN202111176337.4A CN202111176337A CN113789392A CN 113789392 A CN113789392 A CN 113789392A CN 202111176337 A CN202111176337 A CN 202111176337A CN 113789392 A CN113789392 A CN 113789392A
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channel catfish
snp marker
genotype
growth
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CN113789392B (en
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钟立强
王明华
张世勇
姜虎成
陈校辉
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Freshwater Fisheries Research Institute of Jiangsu Province
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Abstract

The invention discloses an SNP marker related to growth of channel catfish and application thereof, wherein the SNP marker is positioned in a muscle regeneration regulation factor of channel catfishMyoGGene sequence SEQ ID NO: 2 at position 391. By utilizing the SNP marker, the channel catfish parent of the 391-th genotype AA is reserved in production, individuals of other genotypes are removed, the channel catfish parent with high growth speed and stable heredity can be quickly selected, the breeding period is shortened, and the breeding process is accelerated.

Description

SNP marker related to growth of channel catfish and application thereof
Technical Field
The invention belongs to the technical field of molecular biology, and particularly relates to an SNP marker related to the growth of channel catfish and application thereof.
Background
Ictalurus punctatus, also known as catfish silurus, is native to North America, has fresh meat, strong disease resistance, rapid growth, easy capture and wide suitable temperature, and after being introduced into China in the 1980 s, the industry develops rapidly, and the annual yield breaks through 30 million tons. However, in the process of rapid development of the channel catfish breeding industry in China, production units do not pay attention to parent seed reserving operation, even large and small ones, so that the genetic diversity of the channel catfish bred is reduced, the growth is slowed down, the specifications are irregular, and the disease resistance is reduced. The growth character directly influences the breeding yield and benefit, so the development of the breeding work of the channel catfish with fast growth and good economic character is very important. The traditional method for breeding fish aiming at growth mainly comprises group breeding and family breeding, and fish with larger individuals and strong bodies are selected as seed reserving parents according to quantitative genetic analysis of phenotypes. The method is time-consuming, labor-consuming and high in workload, and meanwhile, the quality of channel catfish parent is often inaccurately evaluated in phenotype judgment, so that the breeding effect is poor.
With the development of molecular biology and genetics, a plurality of genetic markers, such as AFLP, RAPD, SSR, SNP and other markers, have emerged, wherein the SNP markers are increasingly the first choice genetic markers in genetic breeding research due to wide distribution, suitability for high-throughput automated analysis and stable heredity. SNP markers refer to polymorphisms in genomic DNA sequences caused by single nucleotide changes, mainly caused by single base transitions or transversions. There are relatively few SNPs in the coding region of a gene, because the variation rate in exons is only 1/5 in the surrounding sequence, but it is of great interest in the study of genetic breeding.
Myostatin (MSTN), Insulin-like growth factor-1 (IGF-1) and myogenic regulatory factor (Myogenin, MyoG) all play important roles in regulating animal growth. Therefore, the genes are taken as candidate genes of growth traits, and the correlation analysis of SNP and growth traits is reported in livestock, poultry and aquatic animals. SNP markers related to the gene coding regions and the growth are screened from the channel catfish, channel catfish parents with high growth speed and stable heredity can be quickly selected, the breeding period is shortened, and the breeding process is accelerated.
Disclosure of Invention
The invention aims to provide an SNP marker related to the growth of channel catfish and application thereof, and the SNP marker related to the growth character of the channel catfish MyoG gene coding region is detected for early selection and rapid screening, so that the generation interval is shortened, the selection strength is improved, and the breeding efficiency and accuracy are improved.
The invention also aims to provide a screening method for the rapid-growth channel catfish parent.
The invention adopts the following technical scheme:
an application of a myogenic regulatory factor MyoG gene of channel catfish in parent screening.
As another aspect of the present invention, the present invention provides an SNP marker affecting the growth of channel catfish, wherein: the DNA sequence of the SNP marker is shown as SEQ ID NO: 2, wherein W at position 391 is a base T or A.
Preferably, the SNP marker has the genotype AT the 391 th site of TT, AA and AT.
As another aspect of the invention, the invention provides application of the SNP marker in judging the growth speed of the channel catfish and/or screening the channel catfish which grows quickly.
As another aspect of the present invention, the present invention provides a method for determining the MyoG genotype of ietalurus punetaus, wherein the method detects whether the 391 th base of the SNP marker of the ietalurus punetaus is an AA genotype, wherein the DNA sequence of the SNP marker is as set forth in SEQ ID NO: 2 is shown in the specification; if the strain is AA genotype, the strain can be used as a channel catfish backup parent with high growth speed and stable heredity.
Preferably, the method comprises the following steps of 1) extracting genomic DNA of the channel catfish; 2) and (3) detecting the 391 th genotype of the SNP marker by PCR by taking the extracted genome DNA as a template.
Preferably, in the step 2), the PCR detection is specifically performed by performing PCR amplification on the genomic DNA of the Ictalurus punctatus by using primers F1 and R1 to obtain a PCR product,
F1：5'-TCTTCCCTTCCAGGCTTACA-3'(SEQ ID NO：3)；
R1：5'-AGCAGCCGAGGACCTGTAAT-3'(SEQ ID NO：4)。
the PCR product was detected by agarose gel electrophoresis and then subjected to sequencing analysis.
Preferably, the reaction system of the primary PCR amplification is that the PCR amplification uses a ready-to-use UltraTaq enzyme PCR kit, which comprises 2 xUltraTaq Master Mix 20 ul, genomic DNA 2 ul, and upstream and downstream primersSubstance 2. mu.l, ddH2O14 mu l; the amplification conditions were: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30s, annealing at 59 ℃ for 30s, and extension at 72 ℃ for 60s for 30 cycles; extension at 72 ℃ for 10 min.
Detecting the PCR product by 1% agarose gel electrophoresis, and observing the electrophoresis result by a gel imager. All amplification products were sent to Shanghai Bioengineering, Inc. and sequenced directly using ABI3730 XL sequencer (ABI, USA). And (3) observing a sequencing peak image by using Chromas software, and comparing DNA sequences by combining with ClustalX software to judge the base type of the SNP site. Genotype 391 individuals with AA were significantly higher in body weight and length than AT and TT (P < 0.05).
The invention has the beneficial effects that:
the method greatly improves the screening accuracy of the channel catfish parent, can quickly select the channel catfish backup parent with high growth speed and stable heredity in advance, and can be used for directional culture, thereby reducing the cost. Compared with the traditional method, the method has the advantages of strong purposiveness, direct action effect, simple operation, quick detection, low detection cost and convenience for wide popularization and use.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present application will be further described with reference to specific examples.
Example 1: acquisition of SNP (single nucleotide polymorphism) marker related to growth traits of channel catfish
1. Laboratory animal
The channel catfish samples used in the experiment are all from the experimental base raised by the channel catfish genetic breeding center of the institute of freshwater aquaculture in Jiangsu province. And (3) constructing a G3 generation family of channel catfish core breeding population in 6 months in 2019, and in order to reduce the influence of the environment on the growth of the channel catfish, breeding the seedlings according to the same standard, namely uniform water changing rate, feeding amount, breeding density, oxygen filling amount and water temperature. When the average age of the family is 170 days, measuring the weight and length data of the individual, randomly selecting 6 family 300 fish, collecting tail fin tissues of each fish, soaking in 95% alcohol, and storing at-20 ℃.
2. Primer design
According to the MyoG gene (AY534329.1), the MSTN gene (AF396747.1) and the IGF-1 gene (AH015082.2) of the channel catfish disclosed by the GenBank database, a pair of primers F1/R1, F2/R2 and F3/R3 are respectively designed by using PrimerPremier 5 software, and the sizes of amplification products are 464bp, 550bp and 1776bp respectively.
F1:5'-TGTTGGATTGGTCTGGAGTGG-3'(SEQ ID NO:3)
R1:5'-CGTCTACTCTCCACCTGCTTC-3'(SEQ ID NO:4)
F2:5'-ACGGTGTTCCTGTTACTGC-3'(SEQ ID NO:5)
R2:5'-CACCAGATGTTGCTATGC-3'(SEQ ID NO:6)
F3:5'-AAGACGCACGAGCCAGGATTATT-3'(SEQ ID NO:7)
R3:5'-TGAGGAGCACATTAGCACTTCTGG-3'(SEQ ID NO:8)
3. PCR amplification
The PCR amplification uses a ready-to-use UltraTaq enzyme PCR kit (Shanghai Czeri bioengineering Co., Ltd.), and the reaction system is as follows: 2 × UltraTaq Master Mix 20 μ l, genomic DNA 2 μ l, upstream and downstream primers 2 μ l, ddH2O 14μl。
The MyoG gene amplification conditions were: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30s, annealing at 59 ℃ for 30s, and extension at 72 ℃ for 60s for 30 cycles; extension at 72 ℃ for 10 min. The MSTN gene amplification conditions are as follows: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30s, annealing at 58 ℃ for 30s, and extension at 72 ℃ for 60s for 30 cycles; extension at 72 ℃ for 10 min. The IGF-1 gene amplification conditions were: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30s, annealing at 64 ℃ for 30s, and extension at 72 ℃ for 60s for 30 cycles; extension at 72 ℃ for 10 min.
Detecting the PCR product by 1% agarose gel electrophoresis, and observing the electrophoresis result by a gel imager.
4. Sequencing
The electrophoresis qualified samples were sent to Shanghai Biotechnology engineering Co., Ltd and sequenced directly using ABI3730 XL sequencer (ABI, USA). And (3) observing a sequencing peak image by using Chromas software, and comparing DNA sequences by combining with ClustalX software to judge the base type of the SNP site.
5. SNP typing
MyoG gene found two coding region SNPs in total, g.391T > A and g.489A > G, respectively. Wherein g.489A > G is synonymous mutation, and the coded amino acid is changed after mutation and still is Gln; 391T > A is missense mutation, the coded amino acid is changed, and Ser is mutated into Thr.
Two coding region SNPs, namely g.1859C > G and g.1907C > G/T, are discovered in the MSTN gene together and are synonymous mutations, and the coded amino acid is not changed after mutation and is still Leu.
Five coding region SNPs are found in IGF-1 gene, g.1695g > T, g.1744G > C, g.1747G > A, g.1750G > A and g.1826C > G. Wherein, three mutations of g.1744G > C, g.1747G > A and g.1750G > A are synonymous mutations, and the coded amino acid is not changed after mutation and is still divided into Leu, Thr and Pro; g.1695G > T and g.1826C > G are missense mutations, and the encoded amino acid is changed from Arg to Leu and Gly, respectively.
6. Statistical analysis of frequency of SNP loci
The 3 missense mutation SNP detection methods are utilized to respectively judge the genotypes of the randomly selected 6 families of 300 channel catfishes, and the frequency of SNP loci is counted.
Genotype frequency refers to the ratio of the number of individuals with a certain genotype for a trait to the total number of individuals in a population:
PYY＝NYY/N
wherein, PYYRepresenting the YY genotype frequency, N, of a certain siteYYRepresenting the number of individuals in the population having a YY genotype; n is the total number of individuals detected.
Gene frequency refers to the relative ratio of a certain number of genes in a population to the total number of its alleles:
Py＝(2Nyy+Nyy1+Nyy2+…+Nyyi…+Nyyn)/2N
wherein, PyIndicates allele y frequency, NyyRepresenting the number of individuals in the population with yy genotype, NyyiRepresenting the number of individuals with the yyi genotype in the population, y1-yn are n different multiple alleles of allele y. The statistical results are shown in Table 1.
TABLE 1 genotype and allele frequency of 3 missense mutation SNP sites in Ictalurus punctatus population
Figure BDA0003295773380000051
7. Association analysis of gene effects
The relevance of different genotype individuals of the channel catfish with more comprehensive character records and the growth characters of the channel catfish is remarkably tested (see table 2).
1) The measured body size data mainly comprises: body weight (170 days old) and body length.
2) Correlation analysis a generally linear model: SPSS 20.0 software general linear model GLM (general linear models procedure) was invoked to test the significance of the effect of each genotype on growth traits. The statistical model is as follows:
Yijk＝μ+Gi+Tj+β1P+Eijk
wherein: y isijk: (ii) an individual phenotype record; μ: the overall average; gi: a genotype fixing effect; tj: fixed effect of cultivation time; beta is a1P: linear covariate for initial body weight, Eijk: random error.
From table 2 it can be seen that the individuals with the g.391snp site of MyoG gene of genotype AA are significantly higher in body weight and length than those with the genotypes TT and AT (P < 0.05). And the weight and the body length of the channel catfish with different genotypes at the g.1695 and g.1826SNP sites of the IGF-1 gene have no obvious difference (P is greater than 0.05). Therefore, AA in the single nucleotide polymorphic site (the reference sequence AY534329.1, the 391 th site of the gene) of the MyoG gene of the channel catfish is a dominant genotype and can be used as a DNA marker. Therefore, in the breeding work, an individual with the MyoG gene of the channel catfish as the AA genotype is selected as a channel catfish parent for breeding growth characters.
TABLE 2 Association analysis between different genotypes and growth traits at SNP sites of Ictalurus punctatus
Figure BDA0003295773380000061
Note: the values in the table are mean values ± sem; marked a, b in the same column as the level of significance of difference P < 0.05.
In order to verify the reliability of the excavated 1SNP molecular marker, weight gain measurement and SNP typing based on Sanger sequencing are carried out on the channel catfish breeding population with the market specification, and the fact that the polymorphism of the SNP locus is extremely remarkably associated with the growth traits of the channel catfish (P <0.05) is further verified, so that the SNP marker can be used for selecting the growth traits of the channel catfish.
Example 2: verification of SNP molecular marker related to growth traits of channel catfish
1. Cultivation experiment and sample collection
After the growth traits such as body weight, body length and the like of the G3 generation family of the channel catfish core breeding population are measured for the first time, a PIT electronic chip is injected into the abdominal cavity and is placed into the same 6 mu pond for mixed culture for 18 months, and the market standard is reached. Fishing by netting in 11 months in 2020, randomly selecting 500 tails, measuring weight and body length, and collecting tail fin tissues.
2. Extraction of genomic DNA and SNP typing
Genomic DNA was extracted from the tail fin of the test fish. The PCR amplification uses a ready-to-use UltraTaq enzyme PCR kit (Shanghai Czeri bioengineering Co., Ltd.), and the reaction system is as follows: 2 × UltraTaq Master Mix 20 μ l, genomic DNA 2 μ l, upstream and downstream primers 2 μ l, ddH2O14. mu.l. The amplification primer was F1/R1(SEQ ID NO:3 and SEQ ID NO: 4). The amplification conditions were: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30s, annealing at 59 ℃ for 30s, and extension at 72 ℃ for 60s for 30 cycles; extension at 72 ℃ for 10 min. Detecting the PCR product by 1% agarose gel electrophoresis, and observing the electrophoresis result by a gel imager. Qualified PCR amplification products were Sanger sequenced using ABI3730 sequencer using the same primers as the PCR amplification primers. Sequencing peak maps were read using Chromas software and the SNP sites recorded for each sample (g.391T)>A) The genotype of (a).
3. Correlation analysis of growth traits and SNP
After successfully typing each sample SNP locus (g.391T > A), correlation analysis is carried out on the weight and length traits and the genotypes of the samples. The results are shown in table 3, which indicates that the body weight and body length of AA type individuals are significantly higher than those of TT and AT type individuals (P < 0.05).
TABLE 3 correlation analysis results of growth traits of Ictalurus punctatus and SNP genotypes
Figure BDA0003295773380000071
Note: the values in the table are mean values ± sem; marked a, b in the same column as the level of significance of difference P < 0.05.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Sequence listing
<110> research institute for fresh water and aquatic products in Jiangsu province
<120> SNP marker related to growth of channel catfish and application thereof
<141> 2021-10-09
<160> 8
<170> SIPOSequenceListing 1.0
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Met Glu Leu Phe Glu Thr Asn Pro Tyr Phe Phe Pro Glu Gln Arg Phe
1 5 10 15
Tyr Glu Ser Gly Glu Asn Phe Phe Pro Ser Arg Leu Thr Gly Gly Phe
20 25 30
Asp Gln Gly Gly Tyr Gln Asp Arg Ser Ser Met Val Gly Leu Cys Gly
35 40 45
Asp Gly Arg Leu Leu Ser Ser Asn Val Gly Leu Glu Asp Lys Pro Ser
50 55 60
Pro Ser Ser Thr Leu Ser Leu Ser Leu Ser Pro Asn Gln Glu Gln Glu
65 70 75 80
His Cys Pro Gly Gln Cys Leu Pro Trp Ala Cys Lys Val Cys Lys Arg
85 90 95
Lys Ser Val Ser Met Asp Arg Arg Arg Ala Ala Thr Leu Arg Glu Lys
100 105 110
Arg Arg Leu Lys Lys Val Asn Glu Ala Phe Glu Ala Leu Lys Arg Ser
115 120 125
Thr Leu Ser Asn Pro Asn Gln Arg Leu Pro Lys Val Glu Ile Leu Arg
130 135 140
Ser Ala Ile Gln Tyr Ile Glu Arg Leu Gln Ala Leu Val Ser Ser Leu
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Asn Gln Gln Glu His Glu Gln Thr Gly Leu His Tyr Arg Ser Ser Ala
165 170 175
Ala Gln Arg Val Ser Ser Ser Asn Glu Gln Gly Ser Gly Ser Thr Cys
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Cys Ser Ser Pro Glu Trp Ser Thr Ala Ser Asp His Cys Thr Thr Ala
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Tyr Gly Ser Thr His Glu Asp Leu Leu Asn Glu Asp Ser Ser Glu Gln
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Ala Asn Leu Arg Ser Leu Thr Ser Ile Val Asp Ser Ile Thr Gly Thr
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atggagcttt ttgagaccaa cccatacttc ttccccgagc agcgctttta cgaaagcggt 60
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agctccatgg tggggctgtg tggggacgga aggctgcttt ctagcaatgt aggtctagaa 180
gacaagccat ctccatcctc cacactgtct ctgtcactct cgcccaacca ggagcaagag 240
cactgcccgg ggcaatgcct gccgtgggcc tgcaaggtgt gcaagcgaaa gtcggtaagc 300
atggaccggc gtcgagcggc cacgctacgt gagaagcgca ggctcaagaa ggtaaatgag 360
gcatttgagg cgcttaagcg cagcacactg wcaaacccca accagaggct acccaaggtg 420
gagatcctca gaagtgctat ccagtacatc gagcggctac aggcacttgt cagttcactc 480
aatcagcaag agcatgagca gactggcctg cattacaggt cctcggctgc tcagagggta 540
agttctatca tataggaaag tacattttgg gcaaaaaaat accaggaata tggtagctta 600
tagactgtca tgtcaattat ggatcatttg caaagcaata tatttgttca atttcaacat 660
agagacctgt tcttagctac agattaaaaa atatatagag actatattgc atgtaaagta 720
cacccgattg taacaaaata atctaaaaaa ttgcgcttta acaggtatta tctatattaa 780
acccatgacc atactaacat acactagaat tgtgtacagt tccttataca tatttgctct 840
tggccacacc taaaacacaa accgctccac atcaagttgc gtcagttgta tatgaatggg 900
tcagtattgt gaatgatggg catttgtgct tatgtgtgtg tgtgtaacag gtgtcatcct 960
ccaatgagca gggttcaggc agcacatgct gtagcagccc ggagtggagc actgcatcag 1020
accactgcac cacagcctat ggctccaccc atgaaggtag actcactgaa attaatgaca 1080
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acggtgttcc tgttactgc 19
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caccagatgt tgctatgc 18
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aagacgcacg agccaggatt att 23
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<400> 8
tgaggagcac attagcactt ctgg 24

Claims (9)

1. Flesh regeneration regulating factor of channel catfishMyoGApplication of gene in parent screening.
2. An SNP marker affecting the growth of channel catfish, which is characterized in that: the DNA sequence of the SNP marker is shown as SEQ ID NO: 2, wherein W at position 391 is a base T or A.
3. The SNP marker affecting the growth of Ictalurus punctatus, according to claim 2, wherein: the 391 th genotype of the SNP marker is TT, AA and AT.
4. The application of the SNP marker of claim 2 in judging the growth speed of channel catfish and/or screening the rapidly growing channel catfish.
5. Channel catfishMyoGA method for determining a genotype, characterized by comprising:
detecting whether the 391 th base position of the SNP marker of the channel catfish is an AA genotype or not, wherein the DNA sequence of the SNP marker is shown as SEQ ID NO: 2 is shown in the specification;
if the strain is AA genotype, the strain can be used as a channel catfish backup parent with high growth speed and stable heredity.
6. The method of claim 5, wherein: comprises the following steps of (a) carrying out,
1) extracting genomic DNA of the channel catfish;
2) and (3) detecting the 391 th genotype of the SNP marker by PCR by taking the extracted genome DNA as a template.
7. The method of claim 6, wherein: in step 2), the PCR detection is specifically performed,
carrying out PCR amplification on the genomic DNA of the channel catfish by using primers F1 and R1 to obtain a PCR product,
SEQ ID NO：3，F1：5'-TCTTCCCTTCCAGGCTTACA-3'；
SEQ ID NO：4，R1：5'-AGCAGCCGAGGACCTGTAAT-3'。
and 8, detecting the PCR product by agarose gel electrophoresis, and then carrying out sequencing analysis.
9. The method of claim 7, wherein: the reaction system of the primary PCR amplification is that,
PCR amplification Using Ready-to-use UltraTaq enzyme PCR kit, including 2 × UltraTaq Master Mix 20 μ l, genomic DNA 2 μ l, upstream and downstream primers 2 μ l, ddH2O 14 μl；
The amplification conditions were: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30s, annealing at 59 ℃ for 30s, and extension at 72 ℃ for 60s for 30 cycles; extension at 72 ℃ for 10 min.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105969882A (en) * 2016-06-23 2016-09-28 江苏省淡水水产研究所 Haplotype SNP molecular marker associated with rapid growth of Ictalures punctatus and detection method and application thereof
CN113293218A (en) * 2021-06-21 2021-08-24 江苏省淡水水产研究所 SNP molecular marker for selecting weight gain character of channel catfish and application

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105969882A (en) * 2016-06-23 2016-09-28 江苏省淡水水产研究所 Haplotype SNP molecular marker associated with rapid growth of Ictalures punctatus and detection method and application thereof
CN113293218A (en) * 2021-06-21 2021-08-24 江苏省淡水水产研究所 SNP molecular marker for selecting weight gain character of channel catfish and application

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
GREGORY, D.J.等: "Ictalurus punctatus myogenin gene, complete cds, AY534329.1", 《GENBANK》, pages 1 *

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