CN111926084B - Application of NRIP1 gene as marker in identification of beef quality - Google Patents

Abstract

The invention discloses application of an NRIP1 gene serving as a marker in beef quality identification. Detecting the expression level of the NRIP1 gene in the longus dorsum of the cattle to be detected, if the expression level of the NRIP1 gene in the longus dorsum of the cattle to be detected is lower than the expression level of the NRIP1 gene in the longus dorsum of the cattle to be detected, the beef quality is good, otherwise, the beef quality is bad; the control longus dorsum muscle was derived from beef of good quality. Therefore, the beef quality can be identified by detecting the expression level of the NRIP1 gene in the longus muscle of the dorsum of the cattle. The invention has important application value.

Description

Application of NRIP1 gene as marker in identification of beef quality

Technical Field

The invention belongs to the technical field of biology, and particularly relates to application of an NRIP1 gene serving as a marker in beef quality identification.

Background

There is an increasing awareness of the relationship between diet and health, which increases the interest in the nutritional value of foods. Beef is a common food material, and the quality is important in the beef industry, because it is not only the primary factor of beef consumption, but also the property that the beef industry needs to improve. Factors affecting beef quality mainly include genetic factors, fat content and composition, packaging method, feeding, and the like. With the continuous development of scientific technology, the sequencing technology is continuously fused with the conventional molecular genetics and breeding research, so that the molecular breeding process is quickened, and the breeding of target varieties, such as beef cattle varieties with high-quality beef, is facilitated.

The meat quality character is a complex economic character, a large number of genes participate in the formation process of regulating and controlling the meat quality character, most of researches still stay at the preliminary stage of screening a large number of candidate genes at present, and less systematic and deep researches on gene functions are carried out, so that the functional genes with definite action and clear regulation mechanism are less, the genetic markers of beef quality and the information of related functional genes are very limited, the breeding of good beef breeds and the breeding process of new breeds in China are seriously hindered, and therefore, the research of genetic marker screening, functional gene excavation, gene network regulation and action mechanisms is the key of molecular basis theory and technology which must be broken through in the breeding process of high-quality and high-efficiency beef breeds in China.

Genetic improvement of livestock has undergone from QTL localization to GWAS to current histology. QTL localization refers to the analysis of associations between genetic markers and specific traits throughout the genome to determine the location of chromosomes that affect the major loci of a specific trait. GWAS is a genome-wide search for genetic variations closely linked to complex traits or important economic traits. RNA-seq is a high throughput technique that detects the overall transcription of a tissue of a particular species and can quickly learn about almost all transcript information of that species in a particular state. RNA-seq is a popular technology studied today and has important significance for the study of genetic improvement of beef cattle.

Kazakh cattle are old cattle breeds in Xinjiang, have good meat performance and higher beef quality, and can be rapidly fattened under the condition of grazing in grasslands in summer and autumn. Xinjiang brown cattle is a dairy and meat dual-purpose variety, is obtained by long-term hybridization breeding by taking Kazakhstan cattle as a female parent and Switzerland brown cattle (also Alata Wu Niu and a small amount of Kestelomer cattle) as male parent, and has the characteristics of strong disease resistance, coarse feeding resistance, good grazing performance and strong adaptability. The beef quality of Kazakh cattle is significantly higher than that of Xinjiang brown cattle.

Disclosure of Invention

The invention aims to identify or assist in identifying beef quality.

The invention first protects the use of a substance for detecting the expression level and/or activity of NRIP1 protein, which may be at least one of a 1) to a 4):

a1 Identification or assisted identification of beef quality;

a2 Preparing a product for identification or assisted identification of beef quality;

a3 Screening or assisting in screening cattle with different beef quality;

a4 Preparing a product for screening or assisting in screening cattle having different beef quality.

The invention also protects the use of substances and devices for detecting the expression and/or activity of NRIP1 protein, which may be at least one of a 1) to a 4):

a1 Identification or assisted identification of beef quality;

a2 Preparing a product for identification or assisted identification of beef quality;

a3 Screening or assisting in screening cattle with different beef quality;

a4 Preparing a product for screening or assisting in screening cattle having different beef qualities;

the device may be device a and/or device b;

the device A can comprise a data input device 1, a data recording module 1, a data comparing module 1-1 and a conclusion outputting module 1-1;

the data input device 1 is used for inputting the expression quantity and/or activity value of NRIP1 protein;

the data recording module 1 is used for storing the expression quantity and/or activity value of the NRIP1 protein;

the data comparison module 1-1 is used for comparing the expression quantity and/or activity of the NRIP1 protein in the longus muscle of the cattle dorsum to be tested with the expression quantity and/or activity of the NRIP1 protein in the longus muscle of the cattle dorsum to be compared;

the conclusion output module 1-1 is used for displaying a conclusion, namely if the expression quantity and/or activity of NRIP1 protein in the longest muscle of the beef dorsum to be tested is higher than the expression quantity and/or activity of NRIP1 protein in the longest muscle of the control beef dorsum, the conclusion output module 1-1 displays that the beef quality is poor; if the expression quantity and/or activity of the NRIP1 protein in the longus muscle of the cattle dorsum to be tested is not higher than the expression quantity and/or activity of the NRIP1 protein in the longus muscle of the control cattle dorsum, the conclusion output module 1-1 shows that the beef quality is good;

the control longus dorsum muscle is derived from beef with good beef quality;

the device B can comprise a data input device 1, a data recording module 1, a data comparing module 1-2 and a conclusion outputting module 1-2;

the data comparison module 1-2 is used for comparing the expression quantity and/or activity of NRIP1 proteins in a plurality of bovine dorsum longus muscles;

the conclusion output module 1-2 is used to display the conclusion that the lower the expression level and/or activity of NRIP1 protein in the longus muscle of the back of the cattle, the better the beef quality.

The device A specifically comprises the data input device 1, the data recording module 1, the data comparing module 1-1 and the conclusion outputting module 1-1.

The device B specifically comprises the data input equipment 1, the data recording module 1, the data comparing module 1-2 and the conclusion output module 1-2.

The present invention also protects the use of a substance for detecting the expression level of NRIP1 gene, which may be at least one of a 1) to a 4):

a1 Identification or assisted identification of beef quality;

a2 Preparing a product for identification or assisted identification of beef quality;

a3 Screening or assisting in screening cattle with different beef quality;

a4 Preparing a product for screening or assisting in screening cattle having different beef quality.

The present invention also protects the use of a substance and a device for detecting the expression level of NRIP1 gene, which may be at least one of a 1) to a 4):

a1 Identification or assisted identification of beef quality;

a2 Preparing a product for identification or assisted identification of beef quality;

a3 Screening or assisting in screening cattle with different beef quality;

a4 Preparing a product for screening or assisting in screening cattle having different beef qualities;

the device may be device 1 and/or device 2;

the apparatus 1 may comprise a data input device 2, a data recording module 2, a data comparing module 2-1 and a conclusion outputting module 2-1;

the data input device 2 is used for inputting the expression quantity value of the NRIP1 gene;

the data recording module 2 is used for storing the expression quantity value of the NRIP1 gene;

the data comparison module 2-1 is used for comparing the expression quantity of the NRIP1 gene in the longus muscle of the cattle dorsum to be tested with the expression quantity of the NRIP1 gene in the longus muscle of the control cattle dorsum;

the conclusion output module 2-1 is used for displaying a conclusion, namely if the expression level of the NRIP1 gene in the longus muscle of the beef back to be tested is higher than the expression level of the NRIP1 gene in the longus muscle of the control beef back, the conclusion output module 2-1 displays that the beef quality is poor; if the expression level of the NRIP1 gene in the longus muscle of the beef dorsum to be tested is not higher than the expression level of the NRIP1 gene in the longus muscle of the control beef dorsum, the conclusion output module 2-1 shows that the beef quality is good;

the control longus dorsum muscle is derived from beef with good beef quality;

the apparatus 2 may include a data input device 2, a data recording module 2, a data comparing module 2-2, and a conclusion outputting module 2-2;

the data comparison module 2-2 is used for comparing the expression quantity of the NRIP1 genes in a plurality of bovine dorsum longus muscles;

the conclusion output module 2-2 is used for displaying conclusion that the lower the expression level of the NRIP1 gene in the longus muscle of the beef dorsum is, the better the beef quality is.

The apparatus 1 may in particular be composed of the data input device 2, the data recording module 2, the data comparing module 2-1 and the conclusion outputting module 2-1.

The apparatus 2 may specifically consist of the data input device 2, the data recording module 2, the data comparing module 2-2 and the conclusion outputting module 2-2.

In any of the above applications, the control bovine dorsum longus may be a kazak bovine dorsum longus.

The invention also protects a kit, which can comprise a substance for detecting the expression amount and/or activity of the NRIP1 protein and/or a substance for detecting the expression amount of the NRIP1 gene; the function of the kit may be at least one of a 1) to a 4):

a1 Identification or assisted identification of beef quality;

a2 Preparing a product for identification or assisted identification of beef quality;

a3 Screening or assisting in screening cattle with different beef quality;

a4 Preparing a product for screening or assisting in screening cattle having different beef quality.

The kit may specifically consist of the substance for detecting the expression amount and/or activity of NRIP1 protein and/or the substance for detecting the expression amount of NRIP1 gene.

The invention also protects at least one of D1) to D8):

d1 NRIP1 protein as a marker for identification or assisted identification of beef quality;

d2 Use of NRIP1 protein as a marker for the preparation of a product for the identification or assisted identification of beef quality;

d3 Use of NRIP1 protein as a marker in screening or assisting in screening cattle with different beef quality;

d4 Use of NRIP1 protein as a marker for the preparation of a product for screening or assisting in screening cattle having different beef quality;

d5 Application of NRIP1 gene as a marker in identification or auxiliary identification of beef quality;

d6 Use of NRIP1 gene as a marker for the preparation of a product for identification or assisted identification of beef quality;

d7 Application of NRIP1 gene as a marker in screening or assisting in screening cattle with different beef quality;

d8 NRIP1 gene as a marker for use in the preparation of a product for screening or assisting in screening cattle having different beef qualities.

The invention also provides a method for identifying or assisting in identifying the quality of beef.

The method for identifying or assisting in identifying the beef quality, which is protected by the invention, can be specifically a method Q1), comprises the following steps:

(q 1-1) detecting the expression level and/or activity of NRIP1 protein in the longus muscle of the back of the cow to be tested;

(q 1-2) judging the beef quality according to the expression quantity and/or activity of the NRIP1 protein.

In the method, the principle of judging the beef quality according to the expression quantity and/or activity of the NRIP1 protein is specifically as follows: the lower the expression level and/or activity of NRIP1 protein, the better the beef quality.

The method for identifying or assisting in identifying the beef quality, which is protected by the invention, can be specifically a method Q2), comprises the following steps:

(q 2-1) detecting the expression level of NRIP1 gene in the longus muscle of the dorsum of cattle to be detected;

(q 2-2) determining the beef quality based on the expression level of the NRIP1 gene.

In the method, the principle of judging the beef quality according to the expression quantity of the NRIP1 gene is specifically as follows: the lower the expression level of the NRIP1 gene is, the more excellent the beef quality is.

The invention also provides a method for screening or assisting in screening cattle with different beef quality.

The method for screening or assisting in screening cattle with different beef quality, which is protected by the invention, can be specifically a method R1), comprises the following steps:

(r 1-1) detecting the expression level and/or activity of NRIP1 protein in the longus muscle of the back of the cow to be detected;

(r 1-2) determining the beef quality of the cow based on the expression level and/or activity of the NRIP1 protein.

In the method, the principle of judging the beef quality of the cattle according to the expression quantity and/or activity of the NRIP1 protein is specifically as follows: the lower the expression level and/or activity of NRIP1 protein, the better the beef quality of the cattle.

The method for screening or assisting in screening cattle with different beef quality, which is specifically the method R2), comprises the following steps:

(r 2-1) detecting the expression level of NRIP1 gene in the longus muscle of the dorsum of cattle to be detected;

(r 2-2) determining the beef quality of the cow based on the expression level of the NRIP1 gene.

In the method, the principle of judging the beef quality of the cattle according to the expression quantity of the NRIP1 gene is as follows: the lower the expression level of the NRIP1 gene, the more excellent the beef quality of the cattle.

In any of the above methods, the detection of the expression level of the NRIP1 gene in the longus muscle of the back of the cow to be tested can be specifically achieved by using a substance for detecting the expression level of the NRIP1 gene.

Any of the above-mentioned substances for detecting the expression level of the NRIP1 gene may be specifically a primer pair. The primer pair can be specifically prepared from a primer 1:5'-AGGTACTGCCGTCGACAAAG-3' and primer 2: 5'-CCCTGATACGTGTGTGTGCT-3'.

The nucleotide sequence of any one of the NRIP1 genes is shown as SEQ ID NO. 1.

The amino acid sequence of any one of the NRIP1 proteins (namely the protein encoded by the NRIP1 gene) is shown as SEQ ID NO. 2.

Experiments prove that compared with Kazakh cattle, the relative expression level of the NRIP1 gene in the longus muscle of the dorsum of the cattle in Xinjiang brown cattle is obviously increased. Therefore, the beef quality can be identified by detecting the expression level of the NRIP1 gene in the longus muscle of the dorsum of the cattle, and the lower the expression level of the NRIP1 gene is, the better the beef quality is. The invention has important application value.

Drawings

FIG. 1 is a volcanic plot of normalized quantitative analysis mRNA differentially expressed by longus muscle of dorsum of Kazakh cattle and Xinjiang brown cattle of FPKM.

FIG. 2 shows the FPKM values of the NRIP1 gene in the longus muscle of the dorsum of Kazakh cattle and Xinjiang brown cattle.

FIG. 3 is a graph showing the quantitative detection of the relative expression level of the NRIP1 gene in the longus muscle of the dorsum of Kazakh cattle and Xinjiang brown cattle by real-time fluorescence.

Detailed Description

The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.

The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.

In the following examples, the nucleotide sequence of the NRIP1 gene is shown in SEQ ID NO. 1. The amino acid sequence of the protein coded by the NRIP1 gene (namely NRIP1 protein) is shown as SEQ ID NO. 2.

Example 1 obtaining of markers related to beef quality

1. The longus muscle of the dorsum of 3 Kazakh cattle, about 30 months old, was taken as a sample and named HSK1, HSK2 and HSK3 in order. The longus muscle of the dorsum of 3 Xinjiang brown cattle, 30 months old, was taken as a sample and named XH1, XH2 and XH3 in sequence.

2. Extracting mRNA of samples (HSK 1, HSK2, HSK3, XH1, XH2 and XH 3), and then performing high-throughput sequencing to obtain Raw reads; filtering the data (for removing impurity data such as repeated sequence) to obtain Clean reads; aligning the clear reads with a bovine reference genome (the website is http:// www.ensembl.org/bos_taurus/Info/index.) to obtain Mapped reads; further, alignment efficiency was obtained (alignment efficiency=mapped reads/Clean reads×100%).

The statistical results of clear reads, mapped reads and alignment efficiencies are shown in Table 1.

The results show that the comparison efficiency of the Mapped reads of the 6 samples is more than 93%, and the subsequent experiments can be carried out.

TABLE 1

Sample name	Clean reads	Mapped reads	Alignment efficiency
				HSK1	123553706	115333210	93.35％
HSK2	122717414	114575756	93.37％
				HSK3	128772648	120727626	93.75％
XH1	128617984	120466642	93.66％
				XH2	133627008	125417229	93.86％
XH3	134131790	127137812	94.79％

4. Performing normalized quantitative analysis on the number of the Mapped reads and the transcript length in the step 3 by using FPKM; wherein 3 Kazakh calves are used as a control group, and 3 Xinjiang brown calves are used as an experimental group.

Normalized quantitative analysis of FPKM volcanic diagrams of mRNA differentially expressed by longissimus dorsi of kazakhstan and singapore brown cattle are shown in fig. 1. The results showed that the mRNA differentially expressed in the longus muscle of Kazakh cattle and Xinjiang brown cattle amounted to 929, 471 of which were up-regulated and 458 of which were down-regulated.

According to the sequencing result, the inventor of the invention finds that one gene with larger difference of FPKM values in the longus muscle of the dorsum of the Kazakh cattle and the Xinjiang brown cattle is an NRIP1 gene, and the FPKM value of the NRIP1 gene in the longus muscle of the dorsum of the Xinjiang brown cattle is obviously higher than that in the longus muscle of the dorsum of the Kazakh cattle (figure 2). Thus, the NRIP1 gene may be related to beef quality, and the NRIP1 gene may serve as a marker for beef quality.

Example 2 real-time fluorescent quantitative determination of relative expression level of NRIP1 Gene in the longus muscle of the dorsum of Kazakh cattle and Xinjiang Brown cattle

The experiment was repeated three times to average, each time detecting about 3 Kazakh cattle or Xinjiang brown cattle at 30 months of age, each time repeating the steps as follows:

1. the total RNA of the longus muscle of the dorsum of Kazakh cattle or Xinjiang brown cattle is extracted by adopting a Trizo1 method, then a first-chain cDNA is reversely transcribed, the cDNA is diluted by 50 times by using sterile water as a template, and the relative expression level of an NRIP1 gene (GAPDH gene is taken as an internal reference gene) is detected by adopting a SuperReal PreMix Plus (SYBR Green) kit (product of Tiangen company) through real-time fluorescence quantitative PCR.

Primers for detection of NRIP1 gene were 5'-AGGTACTGCCGTCGACAAAG-3' and 5'-CCCTGATACGTGTGTGTGCT-3'.

Primers for detection of GAPDH gene were 5'-ACATACTCAGCACCAGCATCAC-3' and 5'-ATTCTGGCAAAGTGGACATCG-3'.

The relative expression level of the NRIP1 gene in the longus dorsum muscle of kazakhstan cattle was set to 1, and the relative expression level of the NRIP1 gene in the longus dorsum muscle of singapore cattle was shown in fig. 3 (HSK is kazakh and XH is singapore cattle). The results showed that the relative expression level of NRIP1 gene in the longus muscle of the dorsum of the cattle of singapore was significantly increased compared to the kazakhstan cattle, which was substantially consistent with the previous sequencing results.

The results show that the expression level of the NRIP1 gene is closely related to the beef quality, namely the NRIP1 gene can be used as a marker to identify the beef quality.

The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.

<110> Xinjiang institute of livestock research at the academy of livestock sciences

Application of <120> NRIP1 gene as marker in identification of beef quality

<160> 2

<170> PatentIn version 3.5

<210> 1

<211> 3471

<212> DNA

<213> Artificial sequence

<400> 1

atgactcatg gagaagagct gggctctgat gtgcaccagg attctattgt tctaacttac 60

ctagaaggat tactaatgca tcaggcagca gaggggtcag gtactgccgt cgacaaagcg 120

tctgccgggc gtaatgaaga cgatcagaac tttaacattt ctggtagcgc gtttcccacc 180

tgtcagcgta atggtccagt tctcagcaca cacacgtatc agggatctgg catgctgcac 240

ctcaaaaaag ccagactctt gcagtcttct gaggactgga acgcagcaaa gcggaagagg 300

ctgtctgatt ccatcgtaaa tttaaacgta aagaaggagg ctctgctggc tggcatggct 360

gacagtgcgc ctaaaggcaa acaggacagc aaggtactgg cctctctgct tcagtctttc 420

agctctaggc tgcagactgt tgccctgtca caacaaatca ggcagagcct caaggagcaa 480

ggatacgccc tgagtcataa ttctttaaaa gtggagaaag atttaaggtg ctatggggtt 540

gcatcaagtc acttaaaaac tctgttgaag aaaagcaagg ctaaagatca aaagcccgat 600

accagcatcc ctgatgtcac gaaaaccctt atcagagaca ggttcataga gtcacctcat 660

cacgttgggc agagcggaac aaaggtcgtg agcgaaccct tgtcatgtgc tgcaagatta 720

caggctgtcg caagcatggt ggagaaaagg gctagtcctg ccacttcacc caaacccagt 780

gttgcctgta gccaactagc actacttctc tccagtgaag cccatttaca gcagtattct 840

cgagaacatg ctttaaagac acaaaatgca aatcaggcag caagcgaaag acttgctgcc 900

atggccagat tacaagaaaa tggccagaag gacatgggca gtttccagct ctcaaaaggc 960

atatcaggcc atcttaatgg tcaggcaaga acatcatcaa acaaactaat ggctagcaaa 1020

agtacagcat ttcagaatcc agtgggtatt gtcccttctt cccccaaaaa tgcaggctat 1080

aagaactccc tggaaagaaa caatataaaa caggctgcta ataacagttt gcttttacat 1140

cttcttaaaa gccagaccat acctaagccg atgaacggac acagtcacag tgagagagga 1200

agcatttttg aggagagcag cacacctacg actattgatg actactcaga tcccaatcct 1260

agtttcaccg atgagagcag tggcgatgaa agttcttact ccaactgtgt tcccatagac 1320

ttgtcttgca aacaccggat agaaaaaccc gaacccgacc agcctgtttc tctggataac 1380

ttaactcagt ccttgctaaa cacttgggat ccaaaagtcc ccgaagttga cgtcaaagaa 1440

gatcaagata cctcaaagaa ttctaagcta aattcgcacc agaaagtaac ccttcttcag 1500

ttgctgcttg gccataagaa tgaagaaaac atggaaagaa acggcagccc tcaggaagca 1560

cacagcgatg agacaaagtt cagcacacag aattacaccc ggacgtctgt aatagaaagc 1620

cccagcacca acaggactac tcccgtgagc actcctccat tgcttgcatc caccaaagcc 1680

gactctccca tcaacctttc ccaacactct ctggtcatca aatggaattc cccaccatat 1740

gcctgtggtc cccagcctga aaagccagcg aataccgcct cgaaccacct gatggacctt 1800

acaaaaagca aagaatcgca gggggagaaa gccatccaca atgaaggtgc acaaaactcg 1860

gccacgttca gtgccagtaa actgttacaa aatttagcgc aatgtggaat gcagtcttcc 1920

gtgtcagggg aagagcagag acccagtaaa cagctgctga gtgtaaacac agataaacct 1980

ccaggtatga ttgatagact gaatagccct ctgctagcca ataaaacaag tgcagttgaa 2040

gagaagaaag cattcggcag tcacacaata ggtcctgaac caggactttc tgggtctgaa 2100

atagaaaatc tgcttgaaag gcgcaccgtc ctccagttac tgctgggaaa tcccaacaaa 2160

gggaagactg aaaagaaaga gaagatgccc ttaagagatg agagcactca ggaacataca 2220

gatagagctt taagtgaaca aatattgatg gtgaaaataa aatctgagcc ttgtgatgac 2280

ttgcatccgc atgccacggg cacgcacttg agccatgagg ctccgggagc ccccttctta 2340

gggatggccc ctcccatgca gagaagtgca cctgccttac caacatccga ggacttgaaa 2400

ccagagcctg gctcacctca ggatttttct ttctcaaaga atggtctgct gagtcgattg 2460

ctgagacaaa atcaggacag tcacctggct gatgagctgg acagcagtca cagaaatagt 2520

gaactgacac ttgtagaatc gaagaacctt tgcatggtcc ctaagaaaag gaagctttac 2580

accgagccgt tagaaaaccc ctttaaaaag atgaaaaata acatagtcga tgctgcaaac 2640

agtcacagtg ctccggaggt gctgtacggg tccttgctta accagcaaga gctgaaactt 2700

agcagaagtg atcttgagtt taagcatcct gccagtcatg gttcagccag cgaaagtgaa 2760

cccaggaatt ggaccagaga gagcaaaagc ttcaatgtcc tgaaacagct gcttctctca 2820

gaaaactgtg tgagagattt gtctcagcac aggagtaact ctgtggctga cagtaaaaag 2880

aaaggacaca gaaacagtgt gaccaacagc aagcctgaat tcagcattgc ttctctaaac 2940

ggactgatgt gcggtgccac tcagcccggc agttgcatgg tcagcaggac atttccatac 3000

ccaggtgtcg gaaaggcccc ccggagtcct cctttccctg agcacttggg ctgcacaggg 3060

tctagaccag aagctgggct tgtgaatggg tgttccatgc ccagtgagaa gggacccatt 3120

aagtgggtta tcacagatgt ggacaagaat gagtatgaga aagactctcc gagactgacc 3180

aaaactaacc caatactcta ttacatgctc cagaaaggag gcagttctgt taccagtcga 3240

gaaacacagg acagggacat atggagggag ccttcatctg ctgaaagtat ctcacaggtt 3300

acaatcaaag aagagttact tcctcctgca gaaactaaag cttctttctt taatttaagg 3360

agcacttata atagccatat gggaaataat gcttctcgcc cacacagcgc aaacggagaa 3420

gtttatggac ttctgggaaa catgctaaca ataaaaaagg aatcagaata a 3471

<210> 2

<211> 1156

<212> PRT

<213> Artificial sequence

<400> 2

Met Thr His Gly Glu Glu Leu Gly Ser Asp Val His Gln Asp Ser Ile

1 5 10 15

Val Leu Thr Tyr Leu Glu Gly Leu Leu Met His Gln Ala Ala Glu Gly

20 25 30

Ser Gly Thr Ala Val Asp Lys Ala Ser Ala Gly Arg Asn Glu Asp Asp

35 40 45

Gln Asn Phe Asn Ile Ser Gly Ser Ala Phe Pro Thr Cys Gln Arg Asn

50 55 60

Gly Pro Val Leu Ser Thr His Thr Tyr Gln Gly Ser Gly Met Leu His

65 70 75 80

Leu Lys Lys Ala Arg Leu Leu Gln Ser Ser Glu Asp Trp Asn Ala Ala

85 90 95

Lys Arg Lys Arg Leu Ser Asp Ser Ile Val Asn Leu Asn Val Lys Lys

100 105 110

Glu Ala Leu Leu Ala Gly Met Ala Asp Ser Ala Pro Lys Gly Lys Gln

115 120 125

Asp Ser Lys Val Leu Ala Ser Leu Leu Gln Ser Phe Ser Ser Arg Leu

130 135 140

Gln Thr Val Ala Leu Ser Gln Gln Ile Arg Gln Ser Leu Lys Glu Gln

145 150 155 160

Gly Tyr Ala Leu Ser His Asn Ser Leu Lys Val Glu Lys Asp Leu Arg

165 170 175

Cys Tyr Gly Val Ala Ser Ser His Leu Lys Thr Leu Leu Lys Lys Ser

180 185 190

Lys Ala Lys Asp Gln Lys Pro Asp Thr Ser Ile Pro Asp Val Thr Lys

195 200 205

Thr Leu Ile Arg Asp Arg Phe Ile Glu Ser Pro His His Val Gly Gln

210 215 220

Ser Gly Thr Lys Val Val Ser Glu Pro Leu Ser Cys Ala Ala Arg Leu

225 230 235 240

Gln Ala Val Ala Ser Met Val Glu Lys Arg Ala Ser Pro Ala Thr Ser

245 250 255

Pro Lys Pro Ser Val Ala Cys Ser Gln Leu Ala Leu Leu Leu Ser Ser

260 265 270

Glu Ala His Leu Gln Gln Tyr Ser Arg Glu His Ala Leu Lys Thr Gln

275 280 285

Asn Ala Asn Gln Ala Ala Ser Glu Arg Leu Ala Ala Met Ala Arg Leu

290 295 300

Gln Glu Asn Gly Gln Lys Asp Met Gly Ser Phe Gln Leu Ser Lys Gly

305 310 315 320

Ile Ser Gly His Leu Asn Gly Gln Ala Arg Thr Ser Ser Asn Lys Leu

325 330 335

Met Ala Ser Lys Ser Thr Ala Phe Gln Asn Pro Val Gly Ile Val Pro

340 345 350

Ser Ser Pro Lys Asn Ala Gly Tyr Lys Asn Ser Leu Glu Arg Asn Asn

355 360 365

Ile Lys Gln Ala Ala Asn Asn Ser Leu Leu Leu His Leu Leu Lys Ser

370 375 380

Gln Thr Ile Pro Lys Pro Met Asn Gly His Ser His Ser Glu Arg Gly

385 390 395 400

Ser Ile Phe Glu Glu Ser Ser Thr Pro Thr Thr Ile Asp Asp Tyr Ser

405 410 415

Asp Pro Asn Pro Ser Phe Thr Asp Glu Ser Ser Gly Asp Glu Ser Ser

420 425 430

Tyr Ser Asn Cys Val Pro Ile Asp Leu Ser Cys Lys His Arg Ile Glu

435 440 445

Lys Pro Glu Pro Asp Gln Pro Val Ser Leu Asp Asn Leu Thr Gln Ser

450 455 460

Leu Leu Asn Thr Trp Asp Pro Lys Val Pro Glu Val Asp Val Lys Glu

465 470 475 480

Asp Gln Asp Thr Ser Lys Asn Ser Lys Leu Asn Ser His Gln Lys Val

485 490 495

Thr Leu Leu Gln Leu Leu Leu Gly His Lys Asn Glu Glu Asn Met Glu

500 505 510

Arg Asn Gly Ser Pro Gln Glu Ala His Ser Asp Glu Thr Lys Phe Ser

515 520 525

Thr Gln Asn Tyr Thr Arg Thr Ser Val Ile Glu Ser Pro Ser Thr Asn

530 535 540

Arg Thr Thr Pro Val Ser Thr Pro Pro Leu Leu Ala Ser Thr Lys Ala

545 550 555 560

Asp Ser Pro Ile Asn Leu Ser Gln His Ser Leu Val Ile Lys Trp Asn

565 570 575

Ser Pro Pro Tyr Ala Cys Gly Pro Gln Pro Glu Lys Pro Ala Asn Thr

580 585 590

Ala Ser Asn His Leu Met Asp Leu Thr Lys Ser Lys Glu Ser Gln Gly

595 600 605

Glu Lys Ala Ile His Asn Glu Gly Ala Gln Asn Ser Ala Thr Phe Ser

610 615 620

Ala Ser Lys Leu Leu Gln Asn Leu Ala Gln Cys Gly Met Gln Ser Ser

625 630 635 640

Val Ser Gly Glu Glu Gln Arg Pro Ser Lys Gln Leu Leu Ser Val Asn

645 650 655

Thr Asp Lys Pro Pro Gly Met Ile Asp Arg Leu Asn Ser Pro Leu Leu

660 665 670

Ala Asn Lys Thr Ser Ala Val Glu Glu Lys Lys Ala Phe Gly Ser His

675 680 685

Thr Ile Gly Pro Glu Pro Gly Leu Ser Gly Ser Glu Ile Glu Asn Leu

690 695 700

Leu Glu Arg Arg Thr Val Leu Gln Leu Leu Leu Gly Asn Pro Asn Lys

705 710 715 720

Gly Lys Thr Glu Lys Lys Glu Lys Met Pro Leu Arg Asp Glu Ser Thr

725 730 735

Gln Glu His Thr Asp Arg Ala Leu Ser Glu Gln Ile Leu Met Val Lys

740 745 750

Ile Lys Ser Glu Pro Cys Asp Asp Leu His Pro His Ala Thr Gly Thr

755 760 765

His Leu Ser His Glu Ala Pro Gly Ala Pro Phe Leu Gly Met Ala Pro

770 775 780

Pro Met Gln Arg Ser Ala Pro Ala Leu Pro Thr Ser Glu Asp Leu Lys

785 790 795 800

Pro Glu Pro Gly Ser Pro Gln Asp Phe Ser Phe Ser Lys Asn Gly Leu

805 810 815

Leu Ser Arg Leu Leu Arg Gln Asn Gln Asp Ser His Leu Ala Asp Glu

820 825 830

Leu Asp Ser Ser His Arg Asn Ser Glu Leu Thr Leu Val Glu Ser Lys

835 840 845

Asn Leu Cys Met Val Pro Lys Lys Arg Lys Leu Tyr Thr Glu Pro Leu

850 855 860

Glu Asn Pro Phe Lys Lys Met Lys Asn Asn Ile Val Asp Ala Ala Asn

865 870 875 880

Ser His Ser Ala Pro Glu Val Leu Tyr Gly Ser Leu Leu Asn Gln Gln

885 890 895

Glu Leu Lys Leu Ser Arg Ser Asp Leu Glu Phe Lys His Pro Ala Ser

900 905 910

His Gly Ser Ala Ser Glu Ser Glu Pro Arg Asn Trp Thr Arg Glu Ser

915 920 925

Lys Ser Phe Asn Val Leu Lys Gln Leu Leu Leu Ser Glu Asn Cys Val

930 935 940

Arg Asp Leu Ser Gln His Arg Ser Asn Ser Val Ala Asp Ser Lys Lys

945 950 955 960

Lys Gly His Arg Asn Ser Val Thr Asn Ser Lys Pro Glu Phe Ser Ile

965 970 975

Ala Ser Leu Asn Gly Leu Met Cys Gly Ala Thr Gln Pro Gly Ser Cys

980 985 990

Met Val Ser Arg Thr Phe Pro Tyr Pro Gly Val Gly Lys Ala Pro Arg

995 1000 1005

Ser Pro Pro Phe Pro Glu His Leu Gly Cys Thr Gly Ser Arg Pro

1010 1015 1020

Glu Ala Gly Leu Val Asn Gly Cys Ser Met Pro Ser Glu Lys Gly

1025 1030 1035

Pro Ile Lys Trp Val Ile Thr Asp Val Asp Lys Asn Glu Tyr Glu

1040 1045 1050

Lys Asp Ser Pro Arg Leu Thr Lys Thr Asn Pro Ile Leu Tyr Tyr

1055 1060 1065

Met Leu Gln Lys Gly Gly Ser Ser Val Thr Ser Arg Glu Thr Gln

1070 1075 1080

Asp Arg Asp Ile Trp Arg Glu Pro Ser Ser Ala Glu Ser Ile Ser

1085 1090 1095

Gln Val Thr Ile Lys Glu Glu Leu Leu Pro Pro Ala Glu Thr Lys

1100 1105 1110

Ala Ser Phe Phe Asn Leu Arg Ser Thr Tyr Asn Ser His Met Gly

1115 1120 1125

Asn Asn Ala Ser Arg Pro His Ser Ala Asn Gly Glu Val Tyr Gly

1130 1135 1140

Leu Leu Gly Asn Met Leu Thr Ile Lys Lys Glu Ser Glu

1145 1150 1155

Claims (1)

1. Detection of the longissimus in cattle dorsumNRIP1The application of the expression quantity of the gene in distinguishing Kazakh cattle and Xinjiang brown cattle is characterized in that: the detection of the longus muscle of the back of the cowNRIP1The expression quantity of the gene is realized by real-time quantitative PCR amplification; the primer pair for amplification consists of primer 1:5'-AGGTACTGCCGTCGACAAAG-3' and primer 2: 5'-CCCTGATACGTGTGTGTGCT-3'.