CN113278713B - Gene chip, molecular probe combination, kit and application of sheep multi-angle character - Google Patents

Abstract

The invention discloses a gene chip, a molecular probe combination, a kit and application for analyzing sheep multi-angular character, and relates to the technical field of biology.

Description

Gene chip, molecular probe combination, kit and application of sheep multi-angle character

Technical Field

The invention relates to the technical field of biology, in particular to the technical field of biological detection, and more particularly relates to a sheep multi-angular character molecular probe combination, a gene chip, a kit and application.

Background

Sheep is one of the most important economic animals in the world, variety resources, while multi-angle sheep is one of the precious sheep genetic resources, researchers define sheep with more than two top corners as multi-angle sheep (as shown in figure 1), wherein the four-corner phenotype is abundant, also called four-corner sheep, which is only present in a few old sheep varieties, and many multi-angle sheep varieties are in endangered states, and related breed conservation and breeding work is carried out in all countries, such as establishment of multi-angle sheep protection associations to promote breed conservation, breeding and commercial communication of multi-angle sheep populations. The multi-angle sheep breeds currently in europe and america mainly include Icelandic sheep, Manx Logthan sheep, hebridge sheep and the like. Four multi-angle sheep species are also available in China, including Sishui fur sheep, Alletai sheep, Mongolian sheep, and Baschibai sheep. Due to the inconvenience in production management, the proportion of the multi-angle sheep individuals in the varieties is very low, and some of the multi-angle sheep individuals are endangered to be extinct. On the other hand, due to the high ornamental value of the multi-part sheep, in recent years, the cultivation and basic research of the multi-part sheep breed are more and more paid attention as a non-traditional sheep breed.

However, the phenotype of the sheep's horn is difficult to detect at an early stage and can only be determined until it grows to the age of half an year to 1 year. Indirect determination of the polyhedrosis phenotype is therefore an area of interest to researchers in favor of DNA-based diagnostics. Alderson et al believe that the inheritance of horns is regulated by two loci, and that when both loci are recessively pure, the sheep are polyhedrosis phenotype. Icelandic sheep studies indicate that in the pair of the polyhedric trait and the dihedral trait, the polyhedric is dominant inheritance. Research on two foreign multi-angular sheep breeds, namely Jacob sheet and Navajo-Churro sheet, shows that the associated site of the multi-angular phenotype exists at 132Mb of the 2 # chromosome of sheep. The genetic association analysis of Mongolian sheep, Aletai sheep and Sishui fur sheep in China shows that SNP markers which are obviously related to the sheep multi-angular character exist at 132.6Mb of the sheep chromosome 2. Chinese patent document CN105603092A also found a multi-angle SNP marker located 53449bp downstream of the sheep MTX2 gene, but SNPs capable of determining a phenotype are generally highly complex.

Like the commercialized Sheep SNP chips (Illumina provine SNP50 Beadchip (50K), Illumina sheet HD Genotyping Beadchip (680K) and Illumina provine LD (5K)), more than 54K SNP sites of the Sheep whole genome are covered for genetic breeding, whole genome association analysis, quantitative trait locus localization, gene preference, comparative genomics and other researches, although such many SNP sites are gathered. However, the existing sheep SNP chip is mainly based on western sheep data, lacks of SNP data of combination of Chinese sheep varieties and foreign sheep varieties, still has the problems of insufficient site uniformity, insufficient embodiment of functional sites and regions, extremely low frequency sites of over 10 percent of sites in Chinese sheep populations and the like, so that the design of the SNP chip which is suitable for Chinese sheep populations and can quickly and effectively detect the multiangular characters has very important significance.

Disclosure of Invention

The invention provides a molecular probe combination, a gene chip, a kit and application for analyzing the sheep multi-angle character in order to meet the analysis requirement on the sheep multi-angle character in the current sheep variety research and protection in China.

In order to achieve the technical purpose of the invention, the invention provides the following technical scheme:

in a first aspect, the invention provides an application of 270 SNP site combinations in analyzing sheep multi-angular traits, wherein the physical positions of the 270 SNP site combinations are as follows: at position 132825735 of chr 2, the deoxynucleotide is A or T; the deoxynucleotide of the gene is T or G at the 132825818 th position of chr 2, and the deoxynucleotide of the gene is G or A at the 132825839 th position of chr 2; at position 132826220 of chr 2, the deoxynucleotide is A or C; at position 132826222 of chr 2, the deoxynucleotide is A or G; at position 132826454 of chr 2, the deoxynucleotide is T or G; at position 132826730 of chr 2, the deoxynucleotide is T or C; at position 132826732 of chr 2, the deoxynucleotide is T or C; at position 132826954 of chr 2, the deoxynucleotide is T or C; at position 132828895 of chr 2, the deoxynucleotide is C or G; at position 132830333 of chr 2, the deoxynucleotide is G or T; at position 132830887 of chr 2, the deoxynucleotide is T or A; at position 132833221 of chr 2, the deoxynucleotide is T or C; at position 132833405 of chr 2, the deoxynucleotide is A or G; at position 132833590 of chr 2, the deoxynucleotide is G or C; at position 132833591 of chr 2, the deoxynucleotide is G or T; at position 132833609 of chr 2, the deoxynucleotide is G or A; at position 132833616 of chr 2, the deoxynucleotide of the gene is C or T; at position 132833773 of chr 2, the deoxynucleotide is A or G; at position 132833921 of chr 2, the deoxynucleotide is T or G; at position 132834401 of chr 2, the deoxynucleotide is T or G; at position 132834792 of chr 2, the deoxynucleotide is T or G; at position 132835651 of chr 2, the deoxynucleotide is G or A; at position 132836846 of chr 2, the deoxynucleotide is C or A; at position 132837168 of chr 2, the deoxynucleotide is T or C; at position 132837588 of chr 2, the deoxynucleotide is A or G; at position 132837658 of chr 2, the deoxynucleotide is T or C; at position 132837669 of chr 2, the deoxynucleotide is A or G; at position 132837930 of chr 2, the deoxynucleotide is A or G; at position 132838045 of chr 2, the deoxynucleotide is G or A; at position 132838653 of chr 2, the deoxynucleotide is A or G; at position 132838666 of chr 2, the deoxynucleotide is T or C; at position 132838811 of chr 2, the deoxynucleotide of the gene is C or T; at position 132839013 of chr 2, the deoxynucleotide of the gene is C or T; at position 132839228 of chr 2, the deoxynucleotide is A or T; at position 132839669 of chr 2, the deoxynucleotide is T or G; at position 132839909 of chr 2, the deoxynucleotide of the gene is C or T; at position 132840355 of chr 2, the deoxynucleotide is T or C; at position 132840473 of chr 2, the deoxynucleotide of the gene is C or T; at position 132841002 of chr 2, the deoxynucleotide of the gene is C or T; at position 132841062 of chr 2, the deoxynucleotide is T or C; at position 132841268 of chr 2, the deoxynucleotide is G or T; at position 132841617 of chr 2, the deoxynucleotide is C or A; at position 132843091 of chr 2, the deoxynucleotide of the gene is C or T; at position 132843869 of chr 2, the deoxynucleotide is A or T; at position 132843961 of chr 2, the deoxynucleotide is G or A; at position 132844298 of chr 2, the deoxynucleotide is G or A; at position 132845871 of chr 2, the deoxynucleotide of the gene is C or T; at position 132846379 of chr 2, the deoxynucleotide of the gene is C or T; at position 132846593 of chr 2, the deoxynucleotide is C or A; at position 132847560 of chr 2, the deoxynucleotide is C or G; at position 132848537 of chr 2, the deoxynucleotide is G or A; at position 132848553 of chr 2, the deoxynucleotide of the gene is C or T; at position 132848571 of chr 2, the deoxynucleotide is G or A; at position 132848655 of chr 2, the deoxynucleotide is G or A; at position 132849740 of chr 2, the deoxynucleotide of the gene is C or T; at position 132849948 of chr 2, the deoxynucleotide is G or A; at position 132850299 of chr 2, the deoxynucleotide is G or A; at position 132851592 of chr 2, the deoxynucleotide of the gene is C or T; at position 132851628 of chr 2, the deoxynucleotide is A or G; at position 132851666 of chr 2, the deoxynucleotide is T or C; at position 132851943 of chr 2, the deoxynucleotide is T or C; at position 132852031 of chr 2, the deoxynucleotide is A or G; at position 132852614 of chr 2, the deoxynucleotide of the gene is C or T; at position 132852897 of chr 2, the deoxynucleotide is A or C; at position 132854512 of chr 2, the deoxynucleotide is A or G; at position 132854948 of chr 2, the deoxynucleotide is G or A; at position 132855519 of chr 2, the deoxynucleotide is G or A; at position 132855701 of chr 2, the deoxynucleotide is T or G; at position 132855868 of chr 2, the deoxynucleotide of the gene is C or T; at position 132855969 of chr 2, the deoxynucleotide is C or G; at position 132856384 of chr 2, the deoxynucleotide is C or A; at position 132856707 of chr 2, the deoxynucleotide is T or C; at position 132857538 of chr 2, the deoxynucleotide is T or C; at position 132857775 of chr 2, the deoxynucleotide is G or A; at position 132858280 of chr 2, the deoxynucleotide is G or A; at position 132858747 of chr 2, the deoxynucleotide is C or A; at position 132859330 of chr 2, the deoxynucleotide is G or A; at position 132859435 of chr 2, the deoxynucleotide is C or A; at position 132860032 of chr 2, the deoxynucleotide of the gene is C or T; at position 132860207 of chr 2, the deoxynucleotide is G or C; at position 132860248 of chr 2, the deoxynucleotide is A or C; at position 132861376 of chr 2, the deoxynucleotide of the gene is C or T; at position 132861784 of chr 2, the deoxynucleotide is G or A; at position 132863365 of chr 2, the deoxynucleotide is G or A; at position 132863782 of chr 2, the deoxynucleotide is T or A; at position 132863870 of chr 2, the deoxynucleotide is A or G; at position 132863952 of chr 2, the deoxynucleotide is T or C; at position 132865640 of chr 2, the deoxynucleotide is T or G; at position 132868112 of chr 2, the deoxynucleotide is A or G; at position 132868454 of chr 2, the deoxynucleotide is G or A; at position 132870246 of chr 2, the deoxynucleotide of the gene is C or T; at position 132872192 of chr 2, the deoxynucleotide is T or G; at position 132872449 of chr 2, the deoxynucleotide is G or A; at position 132872527 of chr 2, the deoxynucleotide is A or T; at position 132872748 of chr 2, the deoxynucleotide is A or T; at position 132873251 of chr 2, the deoxynucleotide is A or G; at position 132873521 of chr 2, the deoxynucleotide is A or C; at position 132874646 of chr 2, the deoxynucleotide is A or G; at position 132875222 of chr 2, the deoxynucleotide is A or G; at position 132876030 of chr 2, the deoxynucleotide of the gene is C or T; at position 132876681 of chr 2, the deoxynucleotide of the gene is C or T; at position 132877049 of chr 2, the deoxynucleotide is C or A; at position 132877364 of chr 2, the deoxynucleotide of the gene is C or T; at position 132881057 of chr 2, the deoxynucleotide is T or C; at position 132881098 of chr 2, the deoxynucleotide is T or C; at position 132882535 of chr 2, the deoxynucleotide of the gene is C or T; at position 132882608 of chr 2, the deoxynucleotide is T or C; at position 132882853 of chr 2, the deoxynucleotide is T or C; at position 132883132 of chr 2, the deoxynucleotide of the gene is C or T; at position 132883789 of chr 2, the deoxynucleotide is C or G; at position 132883989 of chr 2, the deoxynucleotide of the gene is C or T; at position 132884643 of chr 2, the deoxynucleotide is G or T; at position 132884690 of chr 2, the deoxynucleotide is T or G; at position 132891838 of chr 2, the deoxynucleotide is T or C; at position 132892087 of chr 2, the deoxynucleotide is C or A; at position 60001530 of chr 2, the deoxynucleotide is G or T; at position 60001673 of chr 2, the deoxynucleotide is A or G; at position 60001691 of chr 2, the deoxynucleotide is G or C; at position 60002923 of chr 2, the deoxynucleotide is G or A; at position 60006808 of chr 2, the deoxynucleotide is A or G; at position 60006908 of chr 2, the deoxynucleotide is C or G; at position 60007631 of chr 2, the deoxynucleotide of the gene is C or T; at position 60007812 of chr 2, the deoxynucleotide is A or G; at position 60012432 of chr 2, the deoxynucleotide is G or A; at position 60012594 of chr 2, the deoxynucleotide of the gene is C or T; at position 60014923 of chr 2, the deoxynucleotide is A or G; at position 60015029 of chr 2, the deoxynucleotide is G or T; at position 60015359 of chr 2, the deoxynucleotide is T or C; at position 60015544 of chr 2, the deoxynucleotide is T or C; at position 60016466 of chr 2, the deoxynucleotide is T or C; at position 60017733 of chr 2, the deoxynucleotide is G or A; at position 60017736 of chr 2, the deoxynucleotide is T or C; at position 60017952 of chr 2, the deoxynucleotide is C or G; at position 60018220 of chr 2, the deoxynucleotide is G or C; at position 60018874 of chr 2, the deoxynucleotide is T or C; at position 60021082 of chr 2, the deoxynucleotide of the gene is C or T; at position 60021298 of chr 2, the deoxynucleotide is T or C; at position 60021417 of chr 2, the deoxynucleotide is G or A; at position 60022459 of chr 2, the deoxynucleotide is T or C; at position 60022896 of chr 2, the deoxynucleotide of the gene is C or T; at position 60023802 of chr 2, the deoxynucleotide is A or G; at position 60024130 of chr 2, the deoxynucleotide is G or A; at position 60024907 of chr 2, the deoxynucleotide is A or G; at position 60025366 of chr 2, the deoxynucleotide of the gene is C or T; at position 60029085 of chr 2, the deoxynucleotide is G or A; at position 60029750 of chr 2, the deoxynucleotide is C or G; at position 60032390 of chr 2, the deoxynucleotide is A or G; at position 60033975 of chr 2, the deoxynucleotide is A or G; at position 60034509 of chr 2, the deoxynucleotide is G or A; at position 60035422 of chr 2, the deoxynucleotide is G or A; at position 60035442 of chr 2, the deoxynucleotide is T or C; at position 60035538 of chr 2, the deoxynucleotide of the gene is C or T; at position 60035773 of chr 2, the deoxynucleotide is A or G; at position 60036154 of chr 2, the deoxynucleotide is T or C; at position 60036201 of chr 2, the deoxynucleotide of the gene is C or T; at position 60036339 of chr 2, the deoxynucleotide is G or A; at position 60036369 of chr 2, the deoxynucleotide is G or A; at position 60036453 of chr 2, the deoxynucleotide is T or G; at position 60037337 of chr 2, the deoxynucleotide is C or A; at position 60037642 of chr 2, the deoxynucleotide is A or G; at position 60037928 of chr 2, the deoxynucleotide of the gene is C or T; at position 60038626 of chr 2, the deoxynucleotide is G or A; at position 60038775 of chr 2, the deoxynucleotide is G or A; at position 60039203 of chr 2, the deoxynucleotide is A or G; at position 60039264 of chr 2, the deoxynucleotide of the gene is C or T; at position 60039289 of chr 2, the deoxynucleotide is G or T; at position 60039979 of chr 2, the deoxynucleotide is G or A; at position 60039983 of chr 2, the deoxynucleotide is T or A; at position 60039984 of chr 2, the deoxynucleotide is T or A; at position 60040077 of chr 2, the deoxynucleotide is G or A; at position 60040285 of chr 2, the deoxynucleotide is G or T; at position 60040356 of chr 2, the deoxynucleotide is G or A; at position 60040370 of chr 2, the deoxynucleotide of the gene is C or T; at position 60040394 of chr 2, the deoxynucleotide is A or G; at position 60040460 of chr 2, the deoxynucleotide is A or G; at position 60040864 of chr 2, the deoxynucleotide is G or A; at position 60041430 of chr 2, the deoxynucleotide is A or G; at position 60041451 of chr 2, the deoxynucleotide of the gene is C or T; at position 60041497 of chr 2, the deoxynucleotide is T or C; at position 60041692 of chr 2, the deoxynucleotide is T or A; at position 60041693 of chr 2, the deoxynucleotide is T or C; at position 60041860 of chr 2, the deoxynucleotide is T or C; at position 60041956 of chr 2, the deoxynucleotide is G or A; at position 60041995 of chr 2, the deoxynucleotide is C or G; at position 60042020 of chr 2, the deoxynucleotide is T or A; at position 60042033 of chr 2, the deoxynucleotide is A or G; at position 60042172 of chr 2, the deoxynucleotide is G or A; at position 60042216 of chr 2, the deoxynucleotide is A or T; at position 60042509 of chr 2, the deoxynucleotide is G or A; at position 60043014 of chr 2, the deoxynucleotide is G or A; at position 60043145 of chr 2, the deoxynucleotide is G or A; at position 60043293 of chr 2, the deoxynucleotide is G or T; at position 60043902 of chr 2, the deoxynucleotide of the gene is C or T; at position 60044053 of chr 2, the deoxynucleotide is G or A; at position 60044127 of chr 2, the deoxynucleotide is G or A; at position 60044264 of chr 2, the deoxynucleotide is C or A; at position 60044277 of chr 2, the deoxynucleotide is A or G; at position 60044651 of chr 2, the deoxynucleotide is C or A; at position 60045018 of chr 2, the deoxynucleotide is C or A; at position 60045047 of chr 2, the deoxynucleotide is G or A; at position 60045167 of chr 2, the deoxynucleotide is A or G; at position 60045177 of chr 2, the deoxynucleotide is G or A; at position 60045321 of chr 2, the deoxynucleotide is G or A; at position 60045359 of chr 2, the deoxynucleotide is A or G; at position 60045460 of chr 2, the deoxynucleotide is T or C; at position 60046216 of chr 2, the deoxynucleotide is G or C; at position 60046371 of chr 2, the deoxynucleotide is G or A; at position 60046394 of chr 2, the deoxynucleotide of the gene is C or T; at position 60046528 of chr 2, the deoxynucleotide is A or G; at position 60046575 of chr 2, the deoxynucleotide is G or A; at position 60046895 of chr 2, the deoxynucleotide is T or C; at position 60047094 of chr 2, the deoxynucleotide is G or A; at position 60047361 of chr 2, the deoxynucleotide is G or A; at position 60047487 of chr 2, the deoxynucleotide of the gene is C or T; at position 60047565 of chr 2, the deoxynucleotide is T or C; at position 60047944 of chr 2, the deoxynucleotide of the gene is C or T; at position 60049078 of chr 2, the deoxynucleotide is G or A; at position 60049161 of chr 2, the deoxynucleotide of the gene is C or T; at position 60049242 of chr 2, the deoxynucleotide of the gene is C or T; at position 60049601 of chr 2, the deoxynucleotide is G or A; at position 60049944 of chr 2, the deoxynucleotide is A or C; at position 49900552 of chr 14, the deoxynucleotide is G or T; at position 49900557 of chr 14, the deoxynucleotide is A or C; at position 49900667 of chr 14, the deoxynucleotide is G or A; at position 49900687 of chr 14, the deoxynucleotide is A or G; at position 49900836 of chr 14, the deoxynucleotide is A or G; at position 49900869 of chr 14, the deoxynucleotide is T or G; at position 49902973 of chr 14, the deoxynucleotide is C or A; at position 49903271 of chr 14, the deoxynucleotide is G or A; at position 49903544 of chr 14, the deoxynucleotide is C or G; at position 49904575 of chr 14, the deoxynucleotide is G or C; at position 49905719 of chr 14, the deoxynucleotide is C or G; at position 49907188 of chr 14, the deoxynucleotide is A or C; at position 49909362 of chr 14, the deoxynucleotide is G or T; at position 49911215 of chr 14, the deoxynucleotide is T or C; at position 49913851 of chr 14, the deoxynucleotide is A or C; at position 49914228 of chr 14, the deoxynucleotide is T or C; at position 49914836 of chr 14, the deoxynucleotide is C or T; at position 49915022 of chr 14, the deoxynucleotide is C or T; at position 49916264 of chr 14, the deoxynucleotide is T or C; at position 49917274 of chr 14, the deoxynucleotide is G or A; at position 49917475 of chr 14, the deoxynucleotide is C or T; at position 49918153 of chr 14, the deoxynucleotide is G or T; at position 49918227 of chr 14, the deoxynucleotide is C or T; at position 49918801 of chr 14, the deoxynucleotide is G or A; at position 49919012 of chr 14, the deoxynucleotide is C or T; at position 49920199 of chr 14, the deoxynucleotide is A or T; at position 49920546 of chr 14, the deoxynucleotide is T or C; at position 49920578 of chr 14, the deoxynucleotide is G or C; at position 49921119 of chr 14, the deoxynucleotide is A or G; at position 49921368 of chr 14, the deoxynucleotide is G or A; at position 49922915 of chr 14, the deoxynucleotide is G or C; at position 49922962 of chr 14, the deoxynucleotide is A or G; at position 49923824 of chr 14, the deoxynucleotide is A or G; at position 49923890 of chr 14, the deoxynucleotide is A or G; at position 49923961 of chr 14, the deoxynucleotide is G or T; at position 49924343 of chr 14, the deoxynucleotide is C or T; at position 49924545 of chr 14, the deoxynucleotide is G or A; at position 49924595 of chr 14, the deoxynucleotide is C or T; at position 49925613 of chr 14, the deoxynucleotide is T or C; at position 49927635 of chr 14, the deoxynucleotide is A or G; at position 49931011 of chr 14, the deoxynucleotide is C or T; at position 49932355 of chr 14, the deoxynucleotide is A or G; at position 49933240 of chr 14, the deoxynucleotide is A or G; at position 49938363 of chr 14, the deoxynucleotide is G or A; at position 49940937 of chr 14, the deoxynucleotide is T or C; at position 49946445 of chr 14, the deoxynucleotide is T or C; at position 49947099 of chr 14, the deoxynucleotide is T or C; at position 49947928 of chr 14, the deoxynucleotide is G or A;

wherein, the physical position information of the sites is determined based on sheep v4.0 genome sequence alignment.

In a second aspect, the invention provides a method for analyzing the sheep multi-angle character, which is characterized in that the 270 SNP site genotypes of the genomic DNA of a sheep to be detected are compared with the 270 SNP site genotypes of the genomic DNA of a control sheep; the 270 SNP sites are the 270 SNP sites described in the first aspect.

In a third aspect, the invention provides a molecular probe combination for analyzing sheep polyhedral character, which detects the site information of the first aspect in a sample to be detected.

In a fourth aspect, the invention provides a gene chip for analyzing the sheep multi-angular character, which is loaded with the molecular probe combination of the third aspect.

In a fifth aspect, the present invention provides a kit for analyzing sheep multi-angular traits, which comprises the molecular probe combination of the third aspect or the gene chip of the fourth aspect.

In a sixth aspect, the invention provides a method for analyzing sheep multi-angular traits, which is used for detecting a sample to be detected by using the molecular probe combination of the third aspect, the gene chip of the fourth aspect or the kit of the fifth aspect.

In a seventh aspect, the present invention provides the molecular probe combination of the third aspect, or the gene chip of the fourth aspect, or the kit of the fifth aspect, having any one of the following uses: (1) the application in evaluation of sheep multi-angular characters; (2) the application in sheep variety identification; (3) the application in tracing sheep varieties; (4) the application in pedigree reconstruction of breeding sheep; (5) the application in sheep germplasm resource protection; (6) the application in sheep germplasm resource improvement; (7) the application in screening sheep varieties; (8) application in sheep breeding.

Has the advantages that:

1. the sheep multi-angle character SNP site combination only consisting of 270 SNP sites is provided based on the research on genetic resources of a plurality of sheep at home and abroad, the SNP site combination provided by the invention can quickly detect the multi-angle character of a sheep individual at an early stage without showing phenotype on the gene level so as to judge the multi-angle character of the sheep, effectively preserve the breed of the multi-angle sheep, shorten the generation interval, accelerate the breeding process and save the cost; in addition, by utilizing the multi-angular character SNP locus combination provided by the invention, the identification and tracing of sheep varieties can be realized from the perspective of multi-angular phenotype, and technical support is provided for germplasm resource protection and germplasm resource improvement.

2. Compared with the existing high-density chip, the probe combination, the gene chip and the kit formed based on the sheep multi-angular character SNP locus provided by the invention have the advantages of small flux, low cost, easier analysis, wide universality and wide market prospect.

Drawings

FIG. 1 is a photograph of a sheep having a multi-angular character;

FIG. 2 is a manhattan diagram of a Sishui fur sheep Vs small tail Han sheep (SSS versus SXW) group;

fig. 3 is a result graph of the significance test performed on the determination result of the population threshold analysis according to the present application.

Detailed Description

The invention is further illustrated by reference to the following detailed description of specific embodiments, which are intended to be illustrative only and not to be construed as limiting the invention. Unless otherwise indicated, the technical means used in the examples are conventional means well known to those skilled in the art, and can be performed with reference to the third edition of the original book "bioinformatics and functional genomics" or related books, and bioinformatics software and products used therein are commercially available. Various procedures and methods not described in detail are conventional methods well known in the art, and the sources of materials used, trade names, and components thereof, if necessary, are indicated at the time of first appearance, and the same reagents used thereafter, if not specifically indicated, are the same as those indicated at the time of first appearance.

In addition, it should be noted that the site combination and application provided by the present invention are completed by the inventors of the present application through hard creative work and optimization work.

The features and advantages described in the site combination section above are also applicable to the molecular probe combination, gene chip, kit and application thereof formed based on the site combination, and are not described herein again.

The sheep multi-angular character referred to in the present invention is divided according to the number of sheep horns, and only two horns are few horns, and more than two horns are multi-angular.

The SNP refers to a Single Nucleotide Polymorphism (Single Nucleotide Polymorphism) and mainly refers to a DNA sequence Polymorphism caused by a variation of a Single Nucleotide on a genome level, wherein the variation of the Single Nucleotide includes a variation caused by a transition, a transversion, an insertion or a deletion of a Single base.

It should be noted that the molecular markers referred to in the present invention are all heritable and detectable DNA sequences or proteins, including but not limited to molecular markers based on molecular hybridization, such as RFLP, MinisatelliteDNA; molecular markers based on PCR technology, such as RAPD, STS, SSR and scarr; DNA labeling based on restriction and PCR techniques; molecular markers based on DNA chip technology, such as SNP; analytical labeling techniques based on the development of EST databases, and the like. The molecular marker provided by the invention can be used for genome mapping, gene positioning research, map-based gene cloning, species genetic relationship, systematic classification and the like.

The probe of the present invention is a nucleic acid sequence (DNA or RNA) having a detection label and a known sequence, which is complementary to the target gene, such as Taqman-MGB probe.

It should be noted that the kit of the present invention is any one of the cassettes conventionally used in the art, which contains reagents for detection or experiment, and is convenient for operators to be able to get rid of the heavy reagent preparation and optimization process. In one embodiment of the present invention, the primer for amplifying the site information provided by the present invention, the molecular marker or probe or gene chip for detecting the site information provided by the present invention, the enzyme and buffer solution for amplification, or the fluorescent marker for detection are also included.

Example 1 acquisition of SNP site combinations of multiangular traits

1. Selection of sheep individuals

In order to achieve a more comprehensive coverage of the domestic and foreign Sheep breeds, the applicant carried out the acquisition of genetic information on 248 Sheep individuals throughout asia, europe, africa and the middle east, including 16 wild Sheep asian moloren breeds, 172 local breeds and 60 breeds, specifically related to the small tailed Sheep, Sishui fur Sheep, big tail han Sheep, hole Sheep, hu Sheep of china jiang, ningxia tan Sheep of china, alexan Sheep of china new jiang, bashme Sheep, dupont Sheep, merino Sheep (fine wool Sheep), merino Sheep (superfine wool Sheep), saffron, zelerian black Sheep, surf Sheep, wagggir Sheep, finland Sheep (filsheep) of finland, weisang Sheep island Sheep (oesnant), teddy Sheep (sheed), sheds (shedullander), soyoho tiger knone, gold island (solhelvet), gotten-down Sheep (gold Sheep), and heaven Sheep (drayagarland), and Sheep (dracawarrio Sheep), and Sheep) Afaham Sheep (Afar Sheep), Nigerl's Muboro Sheep (MbioSheep), Nigeria's Yanka Sheep (Yankasa Sheep), African shorn West African Sheep (West African Dwarf Sheep), Wuda Sheep (Uda Sheep), African Bruna's Jialonka Sheep (Djallonk Sheep), Morse Sheep (Mossi Sheep), Sahler Sheep (Sahalian Sheep), West non-Kernel Sheep (Cameroon Sheep), Irelak Arwashi Sheep (Awassia Sheep), Hadamni Sheep (Hamdani Sheep), Asahi Kanzi Sheep (Makh Sheep), Ashira Sheep Shez-Shez (Shirafay), ganzel Sheep (Ghezel Sheep), Afrika Sheep (Afshii Sheep), Sharl Sheep (Shal Sheep), marqui Sheep (Makui Sheep), mohni Sheep (Moghani Sheep), Karakul Sheep of Pakistan (Karakl Sheep), Asian molar Flon of Iranshenschel.

2. Acquisition of Total SNP set of sheep Whole Gene

A sample of the sheep individual carrying genetic information in step 1, including but not limited to blood, cells, tissue, skin, hair, feces, etc., is collected using methods conventional in the art. Genetic information (such as DNA) in a sample is extracted for high-depth sequencing, the two modes of SAMtools and GATK are compared with a sheep 4.0 reference genome (obtained from NCBI) released in 2015, and a common result obtained by the two modes forms a SNP set, wherein 2836 ten thousand SNP sites are counted and used as a total SNP set of a sheep whole gene.

The genetic information (genetic information) referred to in the present invention means information that an organism replicates the same thing as itself, is transmitted from a parent to a daughter, or is transmitted from a cell to a cell every time each cell divides.

It should be noted that, the extraction of genetic information (e.g., DNA) from a sample for high-depth sequencing may be performed by a biological company, such as watsda gene, illumina, etc., the high-depth sequencing method is a conventional method in the art or a method of the biological company, in an embodiment of the present invention, the average sequencing depth is-25.7 ×, and the high-depth sequencing is performed by using a re-sequencing analysis process.

3. Screening of candidate genes and functional regions thereof

3.1 treatment of sheep genetic information samples of different multiangular traits

Screening sheep varieties with different polygonal characters, grouping corresponding genetic information samples according to the obvious difference of the different sheep varieties on the polygonal characters, and taking Sishui fur sheep Vs small tail sheep (SSS versus SXW) as a group in one embodiment of the invention.

3.2 processing of sheep genetic information of different multiangular traits after grouping

The multi-locus allele frequency difference between each sheep group is swept by XP-CLR, functional regions related to the multi-angular character are scanned (the scanned Manhattan graph is shown in figure 2, wherein the figure 2 is Sishui fur sheep Vs caucasian sheep (SSS versus SXW), meanwhile, functional regions related to the multi-angular character in sheep varieties in each group are excavated by pi ratio (namely pi value), and then the intersection of the two results is taken to screen the functional regions related to the multi-angular character.

Screening genes in the region by referring to published gene research results, finally determining 7 candidate genes PCSK5, HOXD3, HOXD1, HOXD10, HOXD9, HOXD8 and MEGF8 which are related to the multi-angular trait and have quite determined functions, and further determining the corresponding functional regions of the candidate genes by perl scripts.

4. Acquisition of multi-angular character SNP site combination

Searching SNP sites corresponding to the functional region of the candidate gene determined in the step 3 in the total SNP set by utilizing bedtools to obtain a multi-angular character site combination which is related to 7 multi-angular characters of PCSK5, HOXD3, HOXD1, HOXD10, HOXD9, HOXD8 and MEGF8 and only comprises 270 SNP sites.

Example 2 use of the combination of multi-angular SNP sites for the preparation of primer combinations and Probe combinations

The skilled in the art designs a primer according to the sequence information of each site in the multi-angular character SNP site combination provided by the invention, and the designed primer is beneficial to secondary structure evaluation and Tm value evaluation, so that the primer which has good specificity and high sensitivity and can realize the detection purpose under the same reaction condition is finally obtained.

Wherein, the secondary structure evaluation and Tm value evaluation can be performed by any one of the methods commonly used in the art, for example, DNA folding form is used to evaluate the secondary structure, see the description

The Tm value was then evaluated by using the software RaW-Probe.

The methods are all conventional methods, and can be obtained according to the site information in the multi-angular character SNP site combination provided by the application without creative labor, so that the primer obtained according to the multi-angular character SNP site combination provided by the application also belongs to the protection scope of the invention.

Similarly, the invention also provides a probe prepared by utilizing the multi-angle character SNP site combination, such as a tanqman probe.

Example 3 combination of SNP sites for multiangular trait for preparation of Gene chip

The SNP gene chip of the present application is prepared by immobilizing the primer or probe obtained in example 2 on a polymer substrate, such as a nylon membrane, nitrocellulose membrane, plastic, silica gel wafer, micro magnetic bead, etc., or immobilizing the probe on a glass plate, or directly synthesizing the primer or probe obtained in example 2 on a hard surface such as glass, etc., using the same method as the conventional method.

It should be noted that, although one skilled in the art can prepare the SNP gene chip for detecting sheep multi-angular character in any way, and also can prepare the SNP gene chip by entrusted to the bio-corporation, the SNP gene chip prepared based on the multi-angular character SNP site combination provided in the present application is within the scope of the present invention.

Example 4 analysis kit for sheep polyhedrosis traits

The SNP detection kit for the multi-angular character provided by the application comprises primers, probes or gene chips obtained based on the combination of the SNP sites obtained in example 1. According to the type of use, corresponding detection reagents are also included, for example, when the Taqman Probe is obtained based on the SNP site combination obtained in example 1, a buffer, a ligase, AceQUniversal U + Probe Master Mix V2, TaqMan Probe, etc. which are conventionally used in a fluorescent quantitative PCR reaction are also included.

Although the SNP kit for detecting the sheep multi-angular character is configured according to different using modes by a person skilled in the art, the SNP kit for detecting the sheep multi-angular character configured based on the multi-angular character SNP site combination provided by the application belongs to the protection scope of the invention.

Example 5 detection of sheep polyhedrosis trait

The genetic chip designed based on the sheep multi-angular character SNP locus combination provided by the embodiment 1 of the application carries out multi-angular character detection on 21 purchased lambs, simultaneously breeds the lambs to adults, and observes the horn character of the adult sheep, wherein the judgment result of the multi-angular character indicates that the number of the horn is more than 2, the judgment result of the small horn indicates that the number of the horn is only 2, and the genetic detection result and the observation result are shown in a table 1:

TABLE 1 test results

Sample(s)	The result of the detection	Observation results
			H01、H08、H09、H10、H11、H20	Multi-angle	3-6 angles
H02-H07、H12-H19、H21	Few corner	Two corners

As can be seen from the results shown in Table 1, the results of the examination of the gene chip designed using the multi-angular SNP site provided by the present invention were consistent with the results of the observation.

The invention acquires the peripheral blood of a test sheep and extracts DNA in the peripheral blood, utilizes the DNA extracted by a gene chip to analyze, obtains the typing result of each site in each lamb (namely whether each site is homozygote, heterozygote, mutant homozygote or base deletion), calculates the frequency value of the typing result of each site, and compares the frequency value with a population threshold value to obtain a detection result.

The population threshold value in the present application is obtained by analyzing a polygon population and a few corner population, and the method is the same as above.

The judgment results of analyzing the multi-angle population and the few-angle population are subjected to significance test (independent sample ManWhitney U test), the results are shown in figure 3, as can be seen from the results in the figure, P is less than 0.01, the difference is extremely significant, and the judgment results obtained by adopting the method have accuracy and effectiveness.

Industrial applications

The sheep multi-angle character SNP site combination only consisting of 270 SNP sites can be prepared into the sheep multi-angle character SNP site combination and the SNP chip by the technical personnel in the field, the multi-angle character of a sheep individual is evaluated on the genome level, so that the breeding selection is carried out at the early stage, the generation interval is shortened, a large amount of breeding cost is saved, the multi-angle sheep variety protection is facilitated, the multi-angle sheep traceability is facilitated, the sheep genealogy is suggested, and the germplasm resource protection and the germplasm resource improvement are carried out.

The above description is only a preferred example for helping understanding the present invention, and is not intended to limit the present invention, and those skilled in the art can make various changes and modifications to the present invention without departing from the spirit of the present invention, and those skilled in the art should make various changes and modifications to the present invention without departing from the spirit of the present invention.

Claims (7)

1.270 SNP locus combinations are applied to analyzing sheep multi-angular traits, and the physical positions of the 270 SNP locus combinations are as follows: at position 132825735 of chr 2, the deoxynucleotide is A or T; at position 132825818 of chr 2, the deoxynucleotide is T or G; at position 132825839 of chr 2, the deoxynucleotide is G or A; at position 132826220 of chr 2, the deoxynucleotide is A or C; at position 132826222 of chr 2, the deoxynucleotide is A or G; at position 132826454 of chr 2, the deoxynucleotide is T or G; at position 132826730 of chr 2, the deoxynucleotide is T or C; at position 132826732 of chr 2, the deoxynucleotide is T or C; at position 132826954 of chr 2, the deoxynucleotide is T or C; at position 132828895 of chr 2, the deoxynucleotide is C or G; at position 132830333 of chr 2, the deoxynucleotide is G or T; at position 132830887 of chr 2, the deoxynucleotide is T or A; at position 132833221 of chr 2, the deoxynucleotide is T or C; at position 132833405 of chr 2, the deoxynucleotide is A or G; at position 132833590 of chr 2, the deoxynucleotide is G or C; at position 132833591 of chr 2, the deoxynucleotide is G or T; at position 132833609 of chr 2, the deoxynucleotide is G or A; at position 132833616 of chr 2, the deoxynucleotide of the gene is C or T; at position 132833773 of chr 2, the deoxynucleotide is A or G; at position 132833921 of chr 2, the deoxynucleotide is T or G; at position 132834401 of chr 2, the deoxynucleotide is T or G; at position 132834792 of chr 2, the deoxynucleotide is T or G; at position 132835651 of chr 2, the deoxynucleotide is G or A; at position 132836846 of chr 2, the deoxynucleotide is C or A; at position 132837168 of chr 2, the deoxynucleotide is T or C; at position 132837588 of chr 2, the deoxynucleotide is A or G; at position 132837658 of chr 2, the deoxynucleotide is T or C; at position 132837669 of chr 2, the deoxynucleotide is A or G; at position 132837930 of chr 2, the deoxynucleotide is A or G; at position 132838045 of chr 2, the deoxynucleotide is G or A; at position 132838653 of chr 2, the deoxynucleotide is A or G; at position 132838666 of chr 2, the deoxynucleotide is T or C; at position 132838811 of chr 2, the deoxynucleotide of the gene is C or T; at position 132839013 of chr 2, the deoxynucleotide of the gene is C or T; at position 132839228 of chr 2, the deoxynucleotide is A or T; at position 132839669 of chr 2, the deoxynucleotide is T or G; at position 132839909 of chr 2, the deoxynucleotide of the gene is C or T; at position 132840355 of chr 2, the deoxynucleotide is T or C; at position 132840473 of chr 2, the deoxynucleotide of the gene is C or T; at position 132841002 of chr 2, the deoxynucleotide of the gene is C or T; at position 132841062 of chr 2, the deoxynucleotide is T or C; at position 132841268 of chr 2, the deoxynucleotide is G or T; at position 132841617 of chr 2, the deoxynucleotide is C or A; at position 132843091 of chr 2, the deoxynucleotide of the gene is C or T; at position 132843869 of chr 2, the deoxynucleotide is A or T; at position 132843961 of chr 2, the deoxynucleotide is G or A; at position 132844298 of chr 2, the deoxynucleotide is G or A; at position 132845871 of chr 2, the deoxynucleotide of the gene is C or T; at position 132846379 of chr 2, the deoxynucleotide of the gene is C or T; at position 132846593 of chr 2, the deoxynucleotide is C or A; at position 132847560 of chr 2, the deoxynucleotide is C or G; at position 132848537 of chr 2, the deoxynucleotide is G or A; at position 132848553 of chr 2, the deoxynucleotide of the gene is C or T; at position 132848571 of chr 2, the deoxynucleotide is G or A; at position 132848655 of chr 2, the deoxynucleotide is G or A; at position 132849740 of chr 2, the deoxynucleotide of the gene is C or T; at position 132849948 of chr 2, the deoxynucleotide is G or A; at position 132850299 of chr 2, the deoxynucleotide is G or A; at position 132851592 of chr 2, the deoxynucleotide of the gene is C or T; at position 132851628 of chr 2, the deoxynucleotide is A or G; at position 132851666 of chr 2, the deoxynucleotide is T or C; at position 132851943 of chr 2, the deoxynucleotide is T or C; at position 132852031 of chr 2, the deoxynucleotide is A or G; at position 132852614 of chr 2, the deoxynucleotide of the gene is C or T; at position 132852897 of chr 2, the deoxynucleotide is A or C; at position 132854512 of chr 2, the deoxynucleotide is A or G; at position 132854948 of chr 2, the deoxynucleotide is G or A; at position 132855519 of chr 2, the deoxynucleotide is G or A; at position 132855701 of chr 2, the deoxynucleotide is T or G; at position 132855868 of chr 2, the deoxynucleotide of the gene is C or T; at position 132855969 of chr 2, the deoxynucleotide is C or G; at position 132856384 of chr 2, the deoxynucleotide is C or A; at position 132856707 of chr 2, the deoxynucleotide is T or C; at position 132857538 of chr 2, the deoxynucleotide is T or C; at position 132857775 of chr 2, the deoxynucleotide is G or A; at position 132858280 of chr 2, the deoxynucleotide is G or A; at position 132858747 of chr 2, the deoxynucleotide is C or A; at position 132859330 of chr 2, the deoxynucleotide is G or A; at position 132859435 of chr 2, the deoxynucleotide is C or A; at position 132860032 of chr 2, the deoxynucleotide of the gene is C or T; at position 132860207 of chr 2, the deoxynucleotide is G or C; at position 132860248 of chr 2, the deoxynucleotide is A or C; at position 132861376 of chr 2, the deoxynucleotide of the gene is C or T; at position 132861784 of chr 2, the deoxynucleotide is G or A; at position 132863365 of chr 2, the deoxynucleotide is G or A; at position 132863782 of chr 2, the deoxynucleotide is T or A; at position 132863870 of chr 2, the deoxynucleotide is A or G; at position 132863952 of chr 2, the deoxynucleotide is T or C; at position 132865640 of chr 2, the deoxynucleotide is T or G; at position 132868112 of chr 2, the deoxynucleotide is A or G; at position 132868454 of chr 2, the deoxynucleotide is G or A; at position 132870246 of chr 2, the deoxynucleotide of the gene is C or T; at position 132872192 of chr 2, the deoxynucleotide is T or G; at position 132872449 of chr 2, the deoxynucleotide is G or A; at position 132872527 of chr 2, the deoxynucleotide is A or T; at position 132872748 of chr 2, the deoxynucleotide is A or T; at position 132873251 of chr 2, the deoxynucleotide is A or G; at position 132873521 of chr 2, the deoxynucleotide is A or C; at position 132874646 of chr 2, the deoxynucleotide is A or G; at position 132875222 of chr 2, the deoxynucleotide is A or G; at position 132876030 of chr 2, the deoxynucleotide of the gene is C or T; at position 132876681 of chr 2, the deoxynucleotide of the gene is C or T; at position 132877049 of chr 2, the deoxynucleotide is C or A; at position 132877364 of chr 2, the deoxynucleotide of the gene is C or T; at position 132881057 of chr 2, the deoxynucleotide is T or C; at position 132881098 of chr 2, the deoxynucleotide is T or C; at position 132882535 of chr 2, the deoxynucleotide of the gene is C or T; at position 132882608 of chr 2, the deoxynucleotide is T or C; at position 132882853 of chr 2, the deoxynucleotide is T or C; at position 132883132 of chr 2, the deoxynucleotide of the gene is C or T; at position 132883789 of chr 2, the deoxynucleotide is C or G; at position 132883989 of chr 2, the deoxynucleotide of the gene is C or T; at position 132884643 of chr 2, the deoxynucleotide is G or T; at position 132884690 of chr 2, the deoxynucleotide is T or G; at position 132891838 of chr 2, the deoxynucleotide is T or C; at position 132892087 of chr 2, the deoxynucleotide is C or A; at position 60001530 of chr 2, the deoxynucleotide is G or T; at position 60001673 of chr 2, the deoxynucleotide is A or G; at position 60001691 of chr 2, the deoxynucleotide is G or C; at position 60002923 of chr 2, the deoxynucleotide is G or A; at position 60006808 of chr 2, the deoxynucleotide is A or G; at position 60006908 of chr 2, the deoxynucleotide is C or G; at position 60007631 of chr 2, the deoxynucleotide of the gene is C or T; at position 60007812 of chr 2, the deoxynucleotide is A or G; at position 60012432 of chr 2, the deoxynucleotide is G or A; at position 60012594 of chr 2, the deoxynucleotide of the gene is C or T; at position 60014923 of chr 2, the deoxynucleotide is A or G; at position 60015029 of chr 2, the deoxynucleotide is G or T; at position 60015359 of chr 2, the deoxynucleotide is T or C; at position 60015544 of chr 2, the deoxynucleotide is T or C; at position 60016466 of chr 2, the deoxynucleotide is T or C; at position 60017733 of chr 2, the deoxynucleotide is G or A; at position 60017736 of chr 2, the deoxynucleotide is T or C; at position 60017952 of chr 2, the deoxynucleotide is C or G; at position 60018220 of chr 2, the deoxynucleotide is G or C; at position 60018874 of chr 2, the deoxynucleotide is T or C; at position 60021082 of chr 2, the deoxynucleotide of the gene is C or T; at position 60021298 of chr 2, the deoxynucleotide is T or C; at position 60021417 of chr 2, the deoxynucleotide is G or A; at position 60022459 of chr 2, the deoxynucleotide is T or C; at position 60022896 of chr 2, the deoxynucleotide of the gene is C or T; at position 60023802 of chr 2, the deoxynucleotide is A or G; at position 60024130 of chr 2, the deoxynucleotide is G or A; at position 60024907 of chr 2, the deoxynucleotide is A or G; at position 60025366 of chr 2, the deoxynucleotide of the gene is C or T; at position 60029085 of chr 2, the deoxynucleotide is G or A; at position 60029750 of chr 2, the deoxynucleotide is C or G; at position 60032390 of chr 2, the deoxynucleotide is A or G; at position 60033975 of chr 2, the deoxynucleotide is A or G; at position 60034509 of chr 2, the deoxynucleotide is G or A; at position 60035422 of chr 2, the deoxynucleotide is G or A; at position 60035442 of chr 2, the deoxynucleotide is T or C; at position 60035538 of chr 2, the deoxynucleotide of the gene is C or T; at position 60035773 of chr 2, the deoxynucleotide is A or G; at position 60036154 of chr 2, the deoxynucleotide is T or C; at position 60036201 of chr 2, the deoxynucleotide of the gene is C or T; at position 60036339 of chr 2, the deoxynucleotide is G or A; at position 60036369 of chr 2, the deoxynucleotide is G or A; at position 60036453 of chr 2, the deoxynucleotide is T or G; at position 60037337 of chr 2, the deoxynucleotide is C or A; at position 60037642 of chr 2, the deoxynucleotide is A or G; at position 60037928 of chr 2, the deoxynucleotide of the gene is C or T; at position 60038626 of chr 2, the deoxynucleotide is G or A; at position 60038775 of chr 2, the deoxynucleotide is G or A; at position 60039203 of chr 2, the deoxynucleotide is A or G; at position 60039264 of chr 2, the deoxynucleotide of the gene is C or T; at position 60039289 of chr 2, the deoxynucleotide is G or T; at position 60039979 of chr 2, the deoxynucleotide is G or A; at position 60039983 of chr 2, the deoxynucleotide is T or A; at position 60039984 of chr 2, the deoxynucleotide is T or A; at position 60040077 of chr 2, the deoxynucleotide is G or A; at position 60040285 of chr 2, the deoxynucleotide is G or T; at position 60040356 of chr 2, the deoxynucleotide is G or A; at position 60040370 of chr 2, the deoxynucleotide of the gene is C or T; at position 60040394 of chr 2, the deoxynucleotide is A or G; at position 60040460 of chr 2, the deoxynucleotide is A or G; at position 60040864 of chr 2, the deoxynucleotide is G or A; at position 60041430 of chr 2, the deoxynucleotide is A or G; at position 60041451 of chr 2, the deoxynucleotide of the gene is C or T; at position 60041497 of chr 2, the deoxynucleotide is T or C; at position 60041692 of chr 2, the deoxynucleotide is T or A; at position 60041693 of chr 2, the deoxynucleotide is T or C; at position 60041860 of chr 2, the deoxynucleotide is T or C; at position 60041956 of chr 2, the deoxynucleotide is G or A; at position 60041995 of chr 2, the deoxynucleotide is C or G; at position 60042020 of chr 2, the deoxynucleotide is T or A; at position 60042033 of chr 2, the deoxynucleotide is A or G; at position 60042172 of chr 2, the deoxynucleotide is G or A; at position 60042216 of chr 2, the deoxynucleotide is A or T; at position 60042509 of chr 2, the deoxynucleotide is G or A; at position 60043014 of chr 2, the deoxynucleotide is G or A; at position 60043145 of chr 2, the deoxynucleotide is G or A; at position 60043293 of chr 2, the deoxynucleotide is G or T; at position 60043902 of chr 2, the deoxynucleotide of the gene is C or T; at position 60044053 of chr 2, the deoxynucleotide is G or A; at position 60044127 of chr 2, the deoxynucleotide is G or A; at position 60044264 of chr 2, the deoxynucleotide is C or A; at position 60044277 of chr 2, the deoxynucleotide is A or G; at position 60044651 of chr 2, the deoxynucleotide is C or A; at position 60045018 of chr 2, the deoxynucleotide is C or A; at position 60045047 of chr 2, the deoxynucleotide is G or A; at position 60045167 of chr 2, the deoxynucleotide is A or G; at position 60045177 of chr 2, the deoxynucleotide is G or A; at position 60045321 of chr 2, the deoxynucleotide is G or A; at position 60045359 of chr 2, the deoxynucleotide is A or G; at position 60045460 of chr 2, the deoxynucleotide is T or C; at position 60046216 of chr 2, the deoxynucleotide is G or C; at position 60046371 of chr 2, the deoxynucleotide is G or A; at position 60046394 of chr 2, the deoxynucleotide of the gene is C or T; at position 60046528 of chr 2, the deoxynucleotide is A or G; at position 60046575 of chr 2, the deoxynucleotide is G or A; at position 60046895 of chr 2, the deoxynucleotide is T or C; at position 60047094 of chr 2, the deoxynucleotide is G or A; at position 60047361 of chr 2, the deoxynucleotide is G or A; at position 60047487 of chr 2, the deoxynucleotide of the gene is C or T; at position 60047565 of chr 2, the deoxynucleotide is T or C; at position 60047944 of chr 2, the deoxynucleotide of the gene is C or T; at position 60049078 of chr 2, the deoxynucleotide is G or A; at position 60049161 of chr 2, the deoxynucleotide of the gene is C or T; at position 60049242 of chr 2, the deoxynucleotide of the gene is C or T; at position 60049601 of chr 2, the deoxynucleotide is G or A; at position 60049944 of chr 2, the deoxynucleotide is A or C; at position 49900552 of chr 14, the deoxynucleotide is G or T; at position 49900557 of chr 14, the deoxynucleotide is A or C; at position 49900667 of chr 14, the deoxynucleotide is G or A; at position 49900687 of chr 14, the deoxynucleotide is A or G; at position 49900836 of chr 14, the deoxynucleotide is A or G; at position 49900869 of chr 14, the deoxynucleotide is T or G; at position 49902973 of chr 14, the deoxynucleotide is C or A; at position 49903271 of chr 14, the deoxynucleotide is G or A; at position 49903544 of chr 14, the deoxynucleotide is C or G; at position 49904575 of chr 14, the deoxynucleotide is G or C; at position 49905719 of chr 14, the deoxynucleotide is C or G; at position 49907188 of chr 14, the deoxynucleotide is A or C; at position 49909362 of chr 14, the deoxynucleotide is G or T; at position 49911215 of chr 14, the deoxynucleotide is T or C; at position 49913851 of chr 14, the deoxynucleotide is A or C; at position 49914228 of chr 14, the deoxynucleotide is T or C; at position 49914836 of chr 14, the deoxynucleotide is C or T; at position 49915022 of chr 14, the deoxynucleotide is C or T; at position 49916264 of chr 14, the deoxynucleotide is T or C; at position 49917274 of chr 14, the deoxynucleotide is G or A; at position 49917475 of chr 14, the deoxynucleotide is C or T; at position 49918153 of chr 14, the deoxynucleotide is G or T; at position 49918227 of chr 14, the deoxynucleotide is C or T; at position 49918801 of chr 14, the deoxynucleotide is G or A; at position 49919012 of chr 14, the deoxynucleotide is C or T; at position 49920199 of chr 14, the deoxynucleotide is A or T; at position 49920546 of chr 14, the deoxynucleotide is T or C; at position 49920578 of chr 14, the deoxynucleotide is G or C; at position 49921119 of chr 14, the deoxynucleotide is A or G; at position 49921368 of chr 14, the deoxynucleotide is G or A; at position 49922915 of chr 14, the deoxynucleotide is G or C; at position 49922962 of chr 14, the deoxynucleotide is A or G; at position 49923824 of chr 14, the deoxynucleotide is A or G; at position 49923890 of chr 14, the deoxynucleotide is A or G; at position 49923961 of chr 14, the deoxynucleotide is G or T; at position 49924343 of chr 14, the deoxynucleotide is C or T; at position 49924545 of chr 14, the deoxynucleotide is G or A; at position 49924595 of chr 14, the deoxynucleotide is C or T; at position 49925613 of chr 14, the deoxynucleotide is T or C; at position 49927635 of chr 14, the deoxynucleotide is A or G; at position 49931011 of chr 14, the deoxynucleotide is C or T; at position 49932355 of chr 14, the deoxynucleotide is A or G; at position 49933240 of chr 14, the deoxynucleotide is A or G; at position 49938363 of chr 14, the deoxynucleotide is G or A; at position 49940937 of chr 14, the deoxynucleotide is T or C; at position 49946445 of chr 14, the deoxynucleotide is T or C; at position 49947099 of chr 14, the deoxynucleotide is T or C; at position 49947928 of chr 14, the deoxynucleotide is G or A;

wherein, the physical position information of the sites is determined based on sheep v4.0 genome sequence alignment.

2. The method for analyzing the sheep multi-angle character comprises the steps of comparing the 270 SNP site genotypes of the genomic DNA of a sheep to be detected with the 270 SNP site genotypes of the genomic DNA of a control sheep;

wherein the 270 SNP sites are 270 SNP site combinations in the use according to claim 1.

3. The molecular probe combination for analyzing the sheep multi-angular character detects 270 SNP locus information in the application of claim 1 in a sample to be detected.

4. A gene chip for analyzing sheep multi-angular traits, which is loaded with the molecular probe combination of claim 3.

5. A kit for analyzing sheep multi-angular traits, comprising the molecular probe combination of claim 3 or the gene chip of claim 4.

6. A method for analyzing sheep multi-angular traits, which comprises the step of detecting a sample to be detected by using the molecular probe combination as claimed in claim 3, the gene chip as claimed in claim 4 or the kit as claimed in claim 5.

7. The use of the molecular probe combination of claim 3 or the gene chip of claim 4 or the kit of claim 5, for any one of the following:

(1) the application in evaluation of sheep multi-angular characters;

(2) the application in sheep variety identification;

(3) the application in tracing sheep varieties;

(4) the application in pedigree reconstruction of breeding sheep;

(5) the application in sheep germplasm resource protection;

(6) the application in sheep germplasm resource improvement;

(7) the application in screening sheep varieties;

(8) application in sheep breeding.

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