Molecular marker related to sheep height and application thereof

A molecular marker, sheep technology, used in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as changing mRNA stability

Pending Publication Date: 2022-01-11
甘肃润牧生物工程有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, synonymous mutations can alter mRNA stability, splicing regulatory sites, miRNA binding sites, or trans

Method used

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  • Molecular marker related to sheep height and application thereof
  • Molecular marker related to sheep height and application thereof
  • Molecular marker related to sheep height and application thereof

Examples

Experimental program
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Effect test

example 1

[0041] Examples of gene amplification 1CAP2

[0042] (1) Primer Design

[0043] In Hu sheep CAP2 gene DNA (GenBank No. included: NC_040260.1) as a template, Oligo7.0 software design a pair of primers M-F, and M-R, the following primer sequence

[0044] CAP2:

[0045] M-F: 5'-CTCTAGACTGCGTGAGCTTC-3 '(SEQ ID NO: 2),

[0046] M-R: 5'-ACTCTGTGCTCCCAATTCG-3 '(SEQ ID NO: 3)

[0047] (2) CAP2 gene amplification and sequencing

[0048] The total volume of PCR reaction 25μL, wherein the DNA template 1μL, 2 × PCR Master Mix 12.4μL, upstream primer 0.8μL, downstream primer 0.8μL, ddH 2 O 10μL. PCR amplification procedure is: 94 ℃ denaturation 3min, 94 ℃ denaturation 30s, annealed 54.5 deg.] C 30s, 72 extending deg.] C 30s, 35 cycles, and finally extending 10min 72 ℃. PCR reaction products with 1.5% agarose gel electrophoresis showed specific amplification obtained 309bp fragment ( figure 1 . The amplified PCR fragments were sequenced, the sequencing results show that the specific nucleotide ...

example 2

[0051] Establishing genotyping assay method of Example 2

[0052] (1) Design of primer sequence

[0053] For the amplified fragment in Example 1 C / T polymorphism site KASPar primers designed to specifically detect the polymorphic site for said nucleotide sequence of said primer pair is KASPar:

[0054] AlleleC for detecting the forward primer A1:

[0055] CCAGATCAGGGACTGAACCCAT (SEQ ID NO: 4);

[0056] AlleleT for detecting a forward primer A2:

[0057] CAGATCAGGGACTGAACCCAC (SEQ ID NO: 5);

[0058] Universal reverse primer C: TGGTGAGGAATCTGCCTGCTAATG (SEQ ID NO: 6).

[0059] Above primer commissioned Beijing Bioengineering Co. synthesis, KASPar primer were diluted each set of primers to 10μmol / L, and in a volume ratio of 12:12:30 (primer A1: Primer A2: primer C) ratio of a mixture alternate uniform.

[0060] (2) DNA quality control

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Abstract

The invention provides a molecular marker related to sheep height and application thereof. PCR amplification and sequence analysis are carried out on a Hu sheep CAP2 gene, which finds that a T/C polymorphic site exists at the 309 site of an amplified fragment, a KASPar primer is further used for detecting polymorphic sites of 1346 sheep and establishing a least square model, correlation analysis is carried out on the genotype and the height, and finally that the amplified CAP2 gene fragment can serve as the molecular marker related to the sheep height is determined. The molecular marker can be used for selecting CC homozygous sheep to enter a core group so as to improve the height character of the sheep, and economic benefits can be increased.

Description

Technical field [0001] The present invention belongs to the field of molecular labeling preparation, and specifically, the CAP2 gene fragment is specifically involved as molecular markers and their applications which affect the high sheep body. Background technique [0002] Cyclosidase-related protein (CAP) is a highly conserved protein during evolution, composed of 474-551 amino acid residues, and the active effects of the actin cell skeleton (Kosmas et al., 2015). In mammals, CAP1 and CAP2 are two homologs of CAP (Matviw, Yu, & Young, 1992; Swiston, Hubberstey, Yu, & Young, 1995; Yu, Swiston, & Young, 1994). CAP1 acts in endopathy and cell movement (Bertling et al., 2004), and is present in stress fibers and rich muscle enrich (Peche et al., 2007). The similarity between CAP1 and CAP2 is about 76%, and the CAP2 gene of knockout mice will result in a decrease in weight and survival rate (Kosmas et al., 2015). In addition, in the lack of CAP2, the synergistic development of calci...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/124C12Q2600/156C12Q2531/113C12Q2561/101C12Q2537/161C12Q2535/137
Inventor 王维民赵利明李发弟张小雪袁律峰李冲张德印李晓龙赵源张煜坤宋其志林长春徐丹周步博王江荟程江博李文馨
Owner 甘肃润牧生物工程有限责任公司
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