Primer group for identifying sika deer, red deer and hybrid deer, kit and application

A sika deer and primer set technology, applied in the biological field, can solve the problems of inability to identify hybrid deer antler, and achieve the effects of rich alleles, high polymorphism, and strong data sharing

Active Publication Date: 2021-12-24
JIANGHAN UNIVERSITY +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the mitochondria come from the female parent, a series of identification methods based on the COI gene can only be used as the basis for identifying the female parent. For hybrid d...

Method used

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  • Primer group for identifying sika deer, red deer and hybrid deer, kit and application
  • Primer group for identifying sika deer, red deer and hybrid deer, kit and application
  • Primer group for identifying sika deer, red deer and hybrid deer, kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0015] The embodiment of the present invention provides a primer set for identifying sika deer, red deer and hybrid deer. The primer set includes: upstream primers and downstream primers. Specifically, the primer set includes SEQ ID NO: 1 to SEQ ID in the sequence listing NO: 400 primers indicated.

[0016] Taking the publicly released red deer genome sequence GCA_002197005.1 as the reference genome, combined with the genome sequencing data of 100 sika deer and 68 red deer individuals obtained from the NCBI SRA database (acceptance number: PRJNA355630), using Samtools (Version 1.2) and BCFtools (Version:1.2) Identify the SNP sites on the genome, compare and analyze with the NT library of NCBI, and screen the MNP markers according to the following principles:

[0017] (1) The marker sequence is shared by sika deer and red deer, but not in other species;

[0018] (2) There are multiple discontinuous SNP differences in the sequence;

[0019] (3) The length of the marker sequenc...

Embodiment 2

[0039] The embodiment of the present invention provides an application of a MNP marker site, the application includes: the application includes: using the MNP marker site shown in SEQ ID NO: 401 to SEQ ID NO: 600 in the sequence listing for sika deer, horse Genetic difference analysis of deer and hybrid deer and pedigree analysis of sika deer, red deer and hybrid deer.

[0040] 29 sika deer, 32 red deer and 7 hybrid deer F1 generations donated by the Chinese Academy of Inspection and Quarantine using the 200 MNP marker sites and kits provided by the embodiment of the present invention, a total of 68 deer individual DNA were MNP marked Site difference analysis, through cluster analysis, it was found that all samples can be divided into three categories: red deer, sika deer and hybrid deer. figure 2 shown. Among them, 14 samples of red deer and 2 samples of sika deer and hybrid deer were grouped together, which is presumed to be caused by the wrong labeling of the sample sourc...

Embodiment 3

[0041] Example 3 Sika deer, red deer, and hybrid deer individual analysis of sika deer bloodlines

[0042] Excluding the sample sequencing data whose sample name was inconsistent with the classification in Example 2, the MNP marker typing data of the remaining 26 sika deer and 25 red deer were analyzed, and the specific MNP allele types that only existed in sika deer were screened. Obtain 78 MNP marker sites, analyze the number of sika deer specific allele types (T) at these 78 sites in the above samples, calculate the specific allele type ratio of sika deer in each sample, and establish the sika deer pedigree calculation method, namely Sika deer pedigree ratio (M) = number of sika deer specific alleles (T) / total number of 78 MNP marker alleles (N), the higher the M value, the closer the sika deer blood relationship. Utilize this method to carry out sika deer lineage analysis to 65 parts of deer individuals in embodiment 2, analysis result is as follows image 3 shown by im...

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Abstract

The invention discloses a primer group for identifying the sika deer, the red deer and the hybrid deer, a kit and application, and belongs to the technical field of biology. The primer group comprises primers as shown in SEQ ID NO: 1 to SEQ ID NO: 400 in a sequence table. According to the application of MNP marker sites, 200 MNP marker sites shown in Table 1 in the specification are used for genetic difference analysis of the sika deer, the red deer and the hybrid deer and blood line analysis of the sika deer, the red deer and the hybrid deer. The primer group provided by the embodiment of the invention has good distinction degree, reproducibility and accuracy, the kit using the primer group can simultaneously have the advantages, and the detection accuracy can be effectively ensured.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer set, kit and application for identifying sika deer, red deer and hybrid deer. Background technique [0002] Deer antler is a traditional and relatively valuable Chinese medicinal material in my country. It has a significant health care effect on enhancing body functions and eliminating fatigue, and has been widely used in clinical medicine. "Pharmacopoeia of the People's Republic of China" (2015 edition) defines: deer antler base is the unossified and densely hairy young horns of male deer (Cervus Nippon Temminck) or red deer (Cervus elaphus Linnaeus). Current studies have shown that the contents of amino acids, nucleoside components, phospholipid components, cholesterol and polyamines in sika deer antler and red deer antler are different, and the content of nucleoside components in sika deer antler is significantly higher than that of red deer antler. This may be the reaso...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/16C12Q2600/156C12Q2535/122C12Q2537/143
Inventor 张英李论陈颖邢冉冉李甜甜彭海周俊飞高利芬陈利红方治伟
Owner JIANGHAN UNIVERSITY
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