CN106167825A - Microsatellite marker that a kind of yellow catfish growing characteristic is relevant and detection thereof and application - Google Patents

Abstract

The invention discloses a microsatellite marker related to the growth characteristics of the yellow catfish and its detection and application; that is, four microsatellite sites are found on the GH sequence of the yellow catfish, the microsatellite sites are amplified by screening specific primers, and analyzed Its association with growth traits, and find the dominant genotypes that are beneficial to growth traits. The results showed that three loci were significantly or extremely significantly correlated with growth traits, the locus GA was significantly correlated with body weight, body length, body height, and body thickness, and was extremely significantly correlated with total length and caudal peduncle height, among which CD was dominant Genotype; site TCTT has a significant correlation with head length and tail peduncle height, wherein BD is the dominant genotype; site AC has a significant correlation with body thickness and head length, wherein DD is the dominant genotype, using the primers of the present invention The four amplified microsatellite sites all have high genetic diversity and have significant or extremely significant correlations with growth traits, and the invention has very important significance for the practice of molecular breeding.

Description

A Microsatellite Marker Related to the Growth Characteristics of Peeled Catfish and Its Detection and Application

technical field

The invention relates to the technical field of genetic breeding of molecular markers, in particular to a microsatellite marker related to the growth characteristics of yellow catfish and its detection and application.

Background technique

Yellow catfish is a small freshwater economic fish widely cultivated in my country. It is popular among consumers because of its tender meat, rich nutrition, and high meat content. The breeding parents of yellow catfish are mainly wild-caught or cultivated populations for several generations. Artificial breeding, its growth rate, disease resistance and breeding performance are weak, so it is of practical significance to carry out two kinds of breeding work of yellow catfish.

At present, many scholars have carried out a large amount of research on the genetic breeding of yellow catfish, and obtained many achievements. With the development of biotechnology, there are more and more methods for fish genetics and breeding technology. Molecular markers mainly refer to gene fragments that can reflect certain differences in genomes of individual organisms or groups, with high polymorphism and distribution. It has a series of advantages such as wide range and co-dominance, and has a wide range of applications in fish genetic breeding and genetic linkage map construction, phylogenetic relationship identification, and germplasm resource research.

Microsatellite markers (microsatellite), also known as short tandem repeats (short tandem repeats, STRs) or simple repeats (SSRs), are simple repeats evenly distributed in eukaryotic genomes, consisting of 2 to 6 nucleosides Acids are composed of tandem repeats, since the number of repeats of the repeat unit is highly variable and abundant among individuals. Each microsatellite DNA is composed of a core sequence and a flanking sequence. The core sequence is arranged in tandem repeats, and the flanking DNA sequence is located at both ends of the core sequence. A specific part of a chromatin region.

Microsatellite markers are the second generation of molecular markers based on PCR. So far, microsatellite marker technology has been widely used in important economic fishes, and has become a research tool for the study of population genetic diversity, population genetic structure, genetic map construction, and functional genes. An important technique for positioning and QTL analysis. The use of molecular markers to breed high-yield varieties with excellent growth characteristics is one of the important ways to improve economic benefits. Obtaining genes and gene loci related to the growth characteristics of yellow catfish can assist in the breeding of high-yield yellow catfish. Reports on genes and gene loci related to fish growth characteristics.

Contents of the invention

The technical problem to be solved by the present invention is to overcome the above-mentioned defects in the prior art and provide a microsatellite marker related to the growth characteristics of the yellow catfish.

The second object of the present invention is to provide primers for specifically detecting the above-mentioned microsatellite markers.

The third object of the present invention is to provide the application of the above-mentioned microsatellite markers.

The purpose of the present invention is achieved through the following technical solutions:

A primer for amplifying microsatellites related to the growth characteristics of yellow catfish, the nucleotide sequences of the primers are shown in SEQ ID NO: 4-9, wherein the primers SEQ ID NO: 4 and SEQ ID NO: 5 amplify The repeated motif of the added microsatellite fragment is (GA) _n1 , and the value range of n1 is 17-35. High correlation; the repeat motif of the microsatellite fragment amplified by primers SEQ ID NO: 6 and SEQ ID NO: 7 is (TCTT) _n2 , and the value range of n2 is 6-14. The head length and tail peduncle are highly correlated; the repeat motif of the microsatellite fragment amplified by primers SEQ ID NO: 8 and SEQ ID NO: 9 is (AC) _n3 , and the value range of n3 is 25-35. The satellite fragments are related to the body thickness and head length of the yellow catfish.

Preferably, the melting temperature of the microsatellite fragment amplified by primers SEQ ID NO: 4 and SEQ ID NO: 5 is 56° C., and the fragment size is 311 bp; the microsatellite fragment amplified by primers SEQ ID NO: 6 and SEQ ID NO: 7 The melting temperature of the fragment is 61° C., and the fragment size is 272 bp. The melting temperature of the microsatellite fragment amplified by primers SEQ ID NO: 8 and SEQ ID NO: 9 is 56° C., and the fragment size is 220 bp.

The applicant found through research that using the above-mentioned microsatellite primers to amplify samples can specifically amplify microsatellite markers (or called microsatellite loci), and these microsatellite markers are closely related to the growth shape of yellow catfish, so they can be used for Molecular breeding of yellow catfish.

The invention also provides a kit containing the microsatellite primer.

According to the embodiment of the present invention, the full-length traits of the yellow catfish can be effectively determined by detecting the above-mentioned microsatellite markers. Therefore, the present invention also provides the use of the microsatellite primer or the kit in the genetic breeding of the yellow catfish. application.

The present invention also provides a method for detecting the full-length traits of yellow catfish, comprising the following steps:

S1. Extract sample DNA;

S2. Utilize the primers of SEQ ID NO:4～9 described in claim 1 or 2 to amplify the sample DNA, analyze the type and genotype of microsatellite markers in the sample DNA, and determine the full-length traits of the yellow catfish.

The present invention also provides a method for breeding fine yellow catfish, comprising the following steps:

S1. Extract sample DNA;

S2. Utilize the primer amplification sample DNA of claim 1 or 2 SEQ ID NO:4～9, analyze the type and genotype of the microsatellite marker in the sample DNA;

S3. Judgment of results: if the sample DNA contains repetitive motif (GA) _n1 , select the sample with genotype CD as excellent yellow catfish; if the sample DNA contains recombination motif (TCTT) _n2 , select the genotype as The sample of BD is an excellent yellow catfish; if the sample DNA contains repeat motif (AC) _n3 , the sample with genotype DD is selected as an excellent yellow catfish.

Compared with the prior art, the present invention has the following beneficial effects:

The present invention obtains the full-length sequence of the yellow catfish GH gene for the first time, finds 4 microsatellite sites on the GH sequence through the SSRHunter software, and analyzes the correlation between the microsatellite sites and growth traits of the yellow catfish for the first time through the GLM model of the SAS software. The loci with significant differences were tested by variance analysis, and the genotypes of the significantly different loci were compared by Duncan method to find the dominant genotypes that are beneficial to growth traits. The test results are: there are three loci that are significantly or extremely significantly correlated with growth traits, locus GA is significantly correlated with body weight, body length, body height, and body thickness, and is extremely significantly correlated with total length and caudal peduncle height. Among them, CD It is the dominant genotype; site TCTT has a significant correlation with head length and tail peduncle height, among which BD is the dominant genotype; site AC has a significant correlation with body thickness and head length (P<0.05), and DD is the dominant genotype , the microsatellite loci in this experiment have a higher probability of being significantly correlated with growth traits, which may be due to the newly discovered 4 microsatellites located on the GH gene, and their polymorphisms have a direct impact on the growth and development of yellow catfish . The four microsatellite loci found in this study all have high genetic diversity, and the loci GA, TCTT, and AC all have significant or extremely significant correlations with growth traits. The present invention has very important implications for the practice of molecular breeding significance.

Description of drawings

Figure 1 is the result of common amplification PCR, in which, Figure 1 (1) is the electrophoresis result of AT and GA primers at the site; Figure 1 (2) is the electrophoresis result of the primers at the site TCTT and AC; the leftmost band is MarkerDL2000, from the top The sizes of the lower bands are 2000bp, 1000bp, 750bp, 500bp, 250bp, 100bp respectively.

Figure 2 shows the typing results of microsatellite molecular marker PCR products on ABI3730xl; among them, Figure 1 (1) is the AT site typing; 1 (2) is the GA site typing; 1 (3) is the TCTT site Typing; 1 (4) for AC site analysis.

detailed description

The content of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but it should not be construed as a limitation of the present invention. Without departing from the spirit and essence of the present invention, any modifications or replacements made to the methods, steps, and conditions of the present invention fall within the scope of the present invention. Unless otherwise specified, the experimental methods used in the examples are conventional methods and techniques well known to those skilled in the art, and the reagents or materials are obtained through commercial channels.

The present invention adopts the method of homologous cloning to obtain the full length of GH of yellow catfish (sequence shown in SEQ ID NO: 1), a total of 3117bp, 5 exons (respectively called exon 1, exon 1 from the 5' end). exon 2, exon 3, exon 4 and exon 5), 4 introns (from the 5' end are called intron 1, intron 2, intron 3, intron Contain 4), 5'UTR is 165bp, 3'UTR is 485bp.

Example 1 Screening and typing of microsatellite loci

The SSRHunter software was used to search the microsatellite loci in the full-length sequence of the GH gene of the yellow catfish to determine the type, location and length of the microsatellite loci. Primer Premier 5.0 software was used to design upstream and downstream primers at about 150 bp upstream and downstream of the SSRs site. See Table 1 for primer information.

In order to realize the fluorescent semi-automatic detection of microsatellite polymorphisms, it is necessary to label the 5' or 3' ends of common primers with fluorescent modification groups. The PCR product amplified by fluorescent primers is detected by electrophoresis on a 96-well capillary 3730xl DNA analyzer. The fluorescent group will emit colored light under the action of the laser. The system automatically collects fluorescent information of different colors, and finally obtains the PCR product. Data information.

Use the primers in Table 1 to carry out ordinary PCR amplification reactions to find out the best reaction conditions for the primers. The reaction system is shown in Table 2.

PCR reaction program: pre-denaturation at 94 °C for 5 min; 38 cycles (denaturation at 94 °C for 30 s, Tm varies according to different primers, annealing at 51-56 °C for 30 s, extension at 72 °C for 25 s); post-extension at 72 °C for 10 min, 4 ℃ forever. The product was detected by 2% agarose gel electrophoresis. After the electrophoresis, the gel imaging system was used to take pictures and save it; the whole process was protected from light, and the PCR product that could amplify the target band was wrapped in tin foil and sent to Huada Gene Company for further processing. Typing detection.

In this experiment, three kinds of fluorescent modification groups were selected, FAM (green, green) for AT and AC sites, HEX (yellow, yellow) for GA sites, and TAMRA (yellow-orange, orange) for TCTT sites.

The results showed that: 4 microsatellites were detected on the GH gene, and the sites were distributed in intron 1, intron 3, and 3'UTR, all of which belonged to simple SSR, and the core sequences were (AT)n, (GA)n1 , (TCTT) n2, (AC) n3. They are named AT, GA, TCTT, and AC respectively. Before the synthesis of fluorescent primers, PCR amplification is performed with ordinary primers to check the quality of primer amplification, and then fluorescent primers are synthesized, and fluorescent groups are labeled at the 5' end of the upstream (or downstream) primers, as shown in Figure 1 The corresponding values of multiple annealing temperatures can be found for each SSRs site and it is proved that these 4 SSRs sites can be amplified.

Utilize microsatellite PCR products to apply 96-capillary 3730xl DNA analyzer to determine the genotype of SSRs, the specific steps are:

(1) Dilute the PCR amplification product of SSR: Take a 96-well PCR plate, add 10ul of ddH ₂ O to each well, and then add 1ul of the sample to each well (the same is true for multi-tube mixing). The dilution is not fixed and depends on the concentration of each SSR amplification product. Centrifuge slightly to mix.

(2) Prepare internal standard ROX500 mixed solution: take a 1.5ml centrifuge tube, add 1020ul Hi-DiFormamide and 60ul ROX500, vortex and mix well (the ratio of reagent volume for each reaction is Hi-DiFormamide: ROX500 = 8.5 ul: 0.5ul).

(3) Take another new 96-well PCR plate, add 1ul of sample dilution mixture to each well, then add 9ul of ROX500 mixture to each well, and centrifuge slightly to mix.

(4) Sample denaturation, 95°C for 4 minutes, 4°C for 1 minute.

(5) Electrophoresis on the sequencer 3730xl.

(6) Apply the software GeneMapper ID v3.1 to check the reproducibility of the electrophoresis, the analysis results of the fragments and the genotyping results, and finally output the results, as shown in Figure 2.

The small peak next to the main peak is a shadow peak, which is called a shadow band/ghost band in gel electrophoresis, which is caused by the sliding mismatch of the Taq enzyme. The typing results of STRs from top to bottom are:

AT: The fluorescent marker is FAM, and the molecular weight is 252, which is homozygous;

GA: The fluorescent marker is HEX, the molecular weight is 321, and 342 is a heterozygote;

TCTT: The fluorescent marker is TAMRA, and the molecular weight is 286, which is homozygous;

AC: The fluorescent marker is FAM, the molecular weight is 216, and 230 is a heterozygote.

Example 2 SSRs alleles and distribution

There are 160 experimental samples in this embodiment, 3 alleles are detected at site AT, 4 alleles are detected at site TCTT and site AC, and 5 alleles are detected at site GA. The alleles and distribution of each locus are shown in Table 3.

Example 3 Analysis of SSRs genetic growth characteristics

The genetic characteristics analysis of the 4 SSR polymorphic sites in the study is shown in Table 4. The average heterozygosity of the 4 sites is 0.5661, the sites AT and TCTT are moderately polymorphic, and the sites GA and AC are highly polymorphic polymorphism, indicating that the polymorphism of these 4 microsatellites is high.

In this study, SSRs were typed in 96-capillary ABI3730xl, and the general linear model (GLM) in SAS software was used to compare the genotypes of microsatellite loci and the main growth traits (weight, total length, body length, body height, Body thickness, head length, tail peduncle length, and tail peduncle height) were used for correlation analysis, and the sites with significant differences were tested by variance analysis, and Duncan's method was used for multiple comparisons. The results showed that there was no significant difference in growth traits among the genotypes in locus AT (Table 5). In locus GA, the body weight of genotype BE and CD was significantly heavier than genotype AB (P<0.05); the total length of genotype BE, DD, EE was significantly longer than genotype AB (P<0.05), and the genotype CD The body length was significantly longer than genotype AB (P<0. 01); the body length of genotype BE, CD, EE was significantly longer than genotype AB (P<0. 05); the body height of genotype BE and CD was significantly higher than that of genotype AB ( P<0.05); the body thickness of genotype CD was significantly thicker than that of genotype AB and BB (P<0.05); the tail height of genotype BB, BC, BD, CD, DD, EE was significantly higher than that of genotype AB (P<0. 05), and the tail peduncle height of genotype BE was significantly higher than that of genotype AB (P<0. 01) (Table 6 and Table 7).

In locus TCTT, the head length of genotype CC was significantly longer than that of genotype BB (P<0. 05); the tail height of genotype BD was significantly higher than that of genotype AB (P<0. 05) (Table 8 and Table 9). In site AC, the body thickness of genotype CC and DD was significantly thicker than that of genotype AA (P<0.05); the head length of genotype BB, CC, CD, and DD was significantly longer than that of genotype BD (P<0.05) (Table 10 and 11).

Note: Different lowercase letters in the same row indicate significant differences (P<0.05), and different uppercase letters in the same row indicate extremely significant differences (P<0.01).

A total of 4 SSRs loci were found on the GH gene of the yellow catfish. The study showed that the locus AT was not associated with growth traits; the locus GA was significantly associated with body weight, body length, body height, and body thickness (P<0.05), and was associated with the whole population. The length and the height of the caudal peduncle are significantly correlated (P<0.01), and there are eight genotypes in total, among which CD is the dominant genotype; the locus TCTT is significantly correlated with the length of the head and the height of the caudal peduncle (P<0.05), and there are seven genotypes in total Genotypes, among which BD is the dominant genotype; site AC has a significant correlation with body thickness and head length (P<0.05), there are 6 genotypes, of which DD is the dominant genotype.

Comparative example 1

When designing microsatellite primers, two pairs of primers were designed, named as: site-F1/R1, a total of four microsatellite sites AT, GA, TCTT, AC. The red marks are PCR primers for the final amplification, which are fluorescently modified.

When the inventor designed primers based on the 4 microsatellites, he designed many primer pairs, and the primers given below could not specifically amplify microsatellite loci, and could not be used for genotyping.

SEQ ID NO: 10 (AT-F1): 5'-GCTCTGTTACCTGATGAA-3'

SEQ ID NO: 11 (AT-R1): 5'-TGAGTATTGAAGCACTTT-3'

SEQ ID NO: 12 (GA-F2): 5'-GAGGGAAACCTGAGGAAGA-3'

SEQ ID NO: 13 (GA-R2): 5'-GGTGAATGGAGGAGAATAA-3'

SEQ ID NO: 14 (TCTT-F2): 5'-GATCCCTGGATTCCAACT-3'

SEQ ID NO: 15 (TCTT-R2): 5'-AGCACCATCGTGAAACTG-3'

SEQ ID NO: 16 (AC-F1): 5'-AAAATCCACCACTAGACC-3'

SEQ ID NO: 17 (AC-R1): 5'-ACTCACCAACACCACAGA-3'.

SEQUENCE LISTING

<110> South China Agricultural University

<120> A Microsatellite Marker Related to the Growth Characteristics of Peeled catfish and Its Detection and Application

<130>

<160> 17

<170> PatentIn version 3.3

<210> 1

<211> 3117

<212>DNA

<213> GH gene sequence

<400> 1

tgatgttatt gtggaaggtg tgaggtgtct catgtcgatg cattaagcat gtagagatca 60

ggtataaaaa ttggacctgc tagcaggagg gaaatcattg tctgaaagtc ggcgtctgat 120

ttttcagtga aaattcgaac caagtttctt cagagagatt ctaaaatggc tcgaggtaag 180

gctctgacct ggtgttacat tacgactgtg atgttacagt gaggaacttt tattgaygtt 240

tttgttttat aagattacta ttagattttc aaattacttt ctgtataaaa tccaccacta 300

gacctgtttt attttgtgcc taatttcaga attatccttg tagatgtatc ctcacagtac 360

acacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacacat420

actgtatatg catttatataa tgtatgctac tctcctgctg ttcttcctct tccacagggt 480

tagtgttgct ctctgtggtg ttggtgagtt tgttctttag ccaaggcgcg aatttgaga 540

accagaggct cttcaacaac gccgtcattc gtgtgcaaca ccttcatcag ctggctgcca 600

agatgatgga tgactttgta agctgacttt acatcattta cagatgttta gtttactgaa 660

gtatgcttta gctaacatgg ttgctgtgct gcaaatataa aggaggaagc tctgttacct 720

gatgaacgca aacagctcag caagatcttc cccctgagtt tctgcaactc ggactccatc 780

gaagctccgg caggcaaaga cgagacccag aaaagctctg tgagtcacaa aactgcctca 840

agaattcatg cgattttaat atatatatat ttaaagatat ttgttattgt gataacgaat 900

ttggtgattt aatgatcatt agctgtgaca caaatgattt aaaggagcaa tatgtaatat 960

ataaaagtgc ttcaatactc ataaaaaatt gaatcatgtc aaagatgctt taaaatgttc 1020

aaaaaaatgt aataaagtag ttgttttact gtttttacag tttctttgat taaggcttct 1080

acttaattaa ttacccaata aaatatcccc cccccaatat ttaaatgttt cttaaaagaa 1140

ataactaaaa aacattactt tttctgaatt ttttaagagt agccctgcaa tgcaaggtat 1200

ctattttaaa gtttcagtta acatttttct ggcccagaaa caagctccac tgtttttttt 1260

tttttttgta aaaggcagct taagagtcaa acatgtctag ctagtagttg atgttttcat 1320

tgctctctta gcgtgatgaa tattgtgatt tacgtgctac agtctcatgc taaagatcgt 1380

ttataagcaa acatgagcat cggagagaag aatactgctt tgcatctctg tgaactttaa 1440

acgagtctgt tttttaacca ggtgctgaag ctactgcaca cttcctaccg tctgatcgag 1500

tcatgggagt tccccagcaa gaacctcggc aaccctaacc atatctctga gaagctggct 1560

gacctgaaga tgggcatcgg cgtgcttatc gaggtgagca cgagcgccac agatcagaca 1620

ccgagtcact gcaatgtgtg cagggagaac agaagagtta atagaatgtg ttagttaaca 1680

tgtatgagta tgtatttccc gtatggacct gtgtgcacaa aaatgatgtt ttgtacaact 1740

tggatgaaag tgtttttttt gtgtcattcc ccatccatt ctctataact aatacatgct 1800

ctggtgctct gggtggctca gaacttgcga tctaagtaca agacttagca gtacaagaaa 1860

cccgtcagtt cattcagcta tatgcggttt taaaacttaa aactaatcag aatagtagct 1920

atttatttac tgttggtaga atctgaccgt tcattcattc aatggaaaag ttggtaaggt 1980

aataataatg taacttttgt aactgtaagt ttgcagttct gatcaattga aataatttta 2040

atcattaaaa tgtaaatgtt tacaccatat gttgtagtct caggaggata aactcttcat 2100

accttccagc cctattaacg tccccggctc agacccggag ctctgctcct ttacacatga 2160

gtgaaattta aacagtaaca atttatagta ttattagat cattttctcc agtattgtag 2220

tttttatata gcgcaggttg gatttattta actcctacat gttaataaac acaataacca 2280

acaggcgtac ttaaggtggt cgttatttat gatcaattca tgtattgtta ttataaagtg 2340

tttcagttat gcagtatgat aaagtttagt gaaaattgtc tagttttatt gtagcaaggc 2400

ttataaagtc actctgctca ctccataggg atgtctggac ggacagacca gcatggacga 2460

gaacgacgct ctggctccgc cattcgagga tttctaccag accttgagcg agggaaacct 2520

gaggaagagc ttccgtctgc tgtcctgctt caagaaggac atgcacaagg tggagaccta 2580

tctaagcgtg gccaagtgca ggagatccct ggattccaac tgcaccctgt agaatgagag 2640

agagagagag agagagagag agagagag attgattgat tgattgacta gtcacagtct 2700

gggatctctc caggcatgac tatcatcttt ctttctttct ttctttcctt ttgtttcttt 2760

gggtttaatt tctgtcttta tgtatttatt ctcctccatt cacctgttta acagatttct 2820

ccatctcagc aaccttaagg atgtaacata tgggaccagt ttcacgatgg tgctatgcaa 2880

attcagcttt atatcaaata acgaataaga aaatggatta aacgtaatta atttagaaat 2940

ggacgaggac gctcctggct aatgaatgta aaatattcag agggtatatt ttcctatcaa 3000

agtctatact cttctaattt taattatggc ctatttctgt gtgaagcttt gactgtctct 3060

gtgtgaaaat ctctctcatt gcttttttgt ggatttccaa taaacctaat ccctggg 3117

<210> 2

<211> 18

<212>DNA

<213>AT-F

<400> 2

gtttctgcaa ctcggact 18

<210> 3

<211> 18

<212>DNA

<213>AT-R

<400> 3

gcatctttga catgattc 18

<210> 4

<211> 18

<212>DNA

<213>GA-F

<400> 4

ctaccagacc ttgagcga 18

<210> 5

<211> 18

<212>DNA

<213>GA-R

<400> 5

ggtgaatgga ggagaata 18

<210> 6

<211> 19

<212>DNA

<213> TCTT-F

<400> 6

gatccctgga ttccaactg 19

<210> 7

<211> 19

<212>DNA

<213> TCTT-R

<400> 7

tagcaccatc gtgaaactg 19

<210> 8

<211> 17

<212>DNA

<213> AC-F

<400> 8

aatccaccac tagacct 17

<210> 9

<211> 17

<212>DNA

<213>AC-R

<400> 9

ctcaccaaca ccacaga 17

<210> 10

<211> 18

<212>DNA

<213>AT-F1

<400> 10

gctctgttac ctgatgaa 18

<210> 11

<211> 18

<212>DNA

<213>AT-R1

<400> 11

tgagtattga agcacttt 18

<210> 12

<211> 19

<212>DNA

<213>GA-F2

<400> 12

gagggaaacc tgaggaaga 19

<210> 13

<211> 19

<212>DNA

<213>GA-R2

<400> 13

ggtgaatgga ggagaataa 19

<210> 14

<211> 18

<212>DNA

<213> TCTT-F2

<400> 14

gatccctgga ttccaact 18

<210> 15

<211> 18

<212>DNA

<213> TCTT-R2

<400> 15

agcaccatcg tgaaactg 18

<210> 16

<211> 18

<212>DNA

<213> AC-F1

<400> 16

aaaatccacc actagacc 18

<210> 17

<211> 18

<212>DNA

<213> AC-R1

<400> 17

actcaccaac accacaga 18

Claims (6)

1. A primer for amplifying the microsatellites relevant to the growth characteristics of yellow catfish, characterized in that the nucleotide sequence of the primer is as shown in SEQ ID NO: 4～9, wherein the primer SEQ ID NO: 4 The repeat motif of the microsatellite fragment amplified by SEQ ID NO: 5 is (GA) _n1 , and the value range of n1 is 17-35. The microsatellite fragment is related to the body weight, full length, body length, Body height, body thickness, and tail peduncle height are correlated; the repeat motif of the microsatellite fragment amplified by primers SEQ ID NO: 6 and SEQ ID NO: 7 is (TCTT) _n2 , and the value range of n2 is 6-14. Microsatellite fragments are related to head length and tail peduncle height of yellow catfish; the repeat motif of microsatellite fragments amplified by primers SEQ ID NO: 8 and SEQ ID NO: 9 is (AC) _n3 , and the value range of n3 is 25 ~35, the microsatellite fragments are related to the body thickness and head length of the peltatus catfish. 2. the microsatellite primer that is used for the analysis of growth characteristic of peltrow catfish according to claim 1, is characterized in that, the melting temperature of the microsatellite fragment amplified by primers SEQ ID NO: 4 and SEQ ID NO: 5 is 56 ℃ , the fragment size is 311bp; the melting temperature of the microsatellite fragment amplified by primers SEQ ID NO: 6 and SEQ ID NO: 7 is 61°C, the fragment size is 272bp, and the primers SEQ ID NO: 8 and SEQ ID NO: 9 are amplified The melting temperature of the microsatellite fragment is 56°C, and the fragment size is 220bp. 3. A test kit containing the microsatellite primer according to claim 1 or 2. 4. the microsatellite primer described in claim 1 or 2 or the application of kit described in claim 3 in the genetic breeding of yellow catfish. 5. a method for detecting the full-length traits of yellow catfish, is characterized in that, comprises the following steps: S1. Extract sample DNA; S2. Utilize the primers of SEQ ID NO:4～9 described in claim 1 or 2 to amplify the sample DNA, analyze the type and genotype of microsatellite markers in the sample DNA, and determine the full-length traits of the yellow catfish. 6. a method for breeding excellent yellow catfish, is characterized in that, comprises the following steps: S1. Extract sample DNA; S2. Utilize the primer amplification sample DNA of claim 1 or 2 SEQ ID NO:4～9, analyze the type and genotype of the microsatellite marker in the sample DNA; S3. Judgment of results: if the sample DNA contains repetitive motif (GA) _n1 , select the sample with genotype CD as excellent yellow catfish; if the sample DNA contains recombination motif (TCTT) _n2 , select the genotype as The sample of BD is an excellent yellow catfish; if the sample DNA contains repeat motif (AC) _n3 , the sample with genotype DD is selected as an excellent yellow catfish.