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SCAP gene related to lactation of buffalo, and application of same as molecular marker

A molecular marker, buffalo technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problem that there is no buffalo SCAP gene, etc., and achieve high fat-free solid content, high total dry matter content, and high lactose rate Effect

Active Publication Date: 2017-07-21
GUANGXI ZHUANG AUTONOMOUS REGION BUFFALO INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far there is no research report on the buffalo SCAP gene as a molecular marker for buffalo milk production traits

Method used

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  • SCAP gene related to lactation of buffalo, and application of same as molecular marker
  • SCAP gene related to lactation of buffalo, and application of same as molecular marker
  • SCAP gene related to lactation of buffalo, and application of same as molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1: Preparation of buffalo genome total DNA

[0056] The blood buffalo genome total DNA (Tiangen, Beijing) was prepared using a blood genome DNA extraction kit, and the specific operation steps were as follows:

[0057] (1) Take 500 μL of whole blood and place it in a 1.5 mL centrifuge tube, add an equal volume of cell lysate CL, mix by inverting, centrifuge at 12000 rpm for 1 min, absorb the supernatant and leave the precipitate; add an equal volume of cell lysate CL, repeat the process Step once; add 4μL RNAase (100mg / mL), shake for 15sec, and let stand at room temperature for 5min;

[0058] (2) Add 20 μL Proteinase K solution, mix well, add 200 μL buffer GB, fully invert and mix well, place at 56°C for 10 min, invert and mix several times during the period, the solution should become clear;

[0059] (3) Add 200 μL of absolute ethanol and mix thoroughly by inversion; flocculent precipitation may appear at this time;

[0060] (4) Add the above-mentioned sol...

Embodiment 2

[0066] Example 2: Acquisition of SCAP gene molecular markers for buffalo milk production traits

[0067] 1. Primer design: using SEQ NO: 1 as a template, using Primer 5.0 primer design software, tested by Oligo software, determined the forward primer for amplifying the buffalo SCAP gene, and entrusted Dalian Bao Biotechnology Co., Ltd. to synthesize it. The DNA sequences of the forward and reverse primers used to amplify the gene are shown in SEQ ID NO: 2-5.

[0068] 2. PCR amplification reaction

[0069] Genomic DNA samples from 50 buffaloes were randomly selected for pooling as templates for PCR reactions.

[0070] (1) PCR reaction: The total reaction volume is 40 μL, including 1 μL of buffalo DNA pool template, 20 μL of 10×LA Taq Mix, 2 μL of primer mixture (10 mM each) and ddH 2 O 17 μL.

[0071] (2) PCR reaction program: 94°C / 3min; 94°C / 30sec, 60°C / 30sec, 72°C / 1.5min, 30 cycles; 72°C for 10min; 4°C storage

[0072] 3. PCR product sequencing, analysis and molecular mar...

Embodiment 3

[0088] Example 3: Gene SCAP associated with buffalo lactation and its application as a molecular marker

[0089] The breed of the test buffalo herd was a hybrid buffalo herd. The association analysis between different genotypes of SCAP genes A1717600G, G1718168A, G1743088A, T1753656C, G1759116A, C1759142T, G1760740A, A1762368G, T1766036C loci and milk production traits was carried out in buffalo herds.

[0090] The results of association analysis of buffalo SCAP genes A1717600G, G1718168A, G1743088A, T1753656C, G1759116A, C1759142T, G1760740A, A1762368G, T1766036C are shown in Table 2. From Table 2, it can be concluded that the genotype is significantly correlated with the milk production traits of buffalo (P<0.05).

[0091] For locus A1717600G, the milk yield of buffalo individuals carrying GA genotype was higher than that of individuals with AA and GG genotypes.

[0092] For the G1760740A locus, the milk yield of buffalo individuals carrying the GG genotype was higher than...

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Abstract

The invention provides a SCAP gene related to lactation of a buffalo, and application of the same as a molecular marker, belonging to the field of molecular markers for livestock. The nucleotide sequence of the gene is as shown in SEQ ID No. 1, wherein 9 mutant bases are respectively located at site 1717600 bp, site 1718168 bp, site 1743088 bp, site 1753656 bp, site 1759116 bp, site 1759142 bp, site 1760740 bp, site 1762368 bp and site 1766036 bp, and the 9 SNP sites are all substantially related to the milk production properties of the buffalo. The invention discloses the sequences of upstream and downstream primers needed by all the base mutation sites in SEQ ID No. 1 in a detection sequence table. Application of the SCAP gene is beneficial for improving the milk production performance of the buffalo and provides technical support and theoretical foundation for breeding of milk buffalos with excellent milk production performance.

Description

[0001] 【Technical field】 [0002] The invention relates to the technical field, in particular to the field of animal molecular markers. It specifically relates to the gene SCAP related to buffalo lactation and its application as a molecular marker. [0003] 【Background technique】 [0004] Buffalo is a unique breeding stock resource in southern my country, but its low milk yield and reproductive performance have become two major scientific problems affecting its industrial development. Obviously, the use of advanced molecular marker technology in the selection of high-quality milk buffalo germplasm resources will effectively improve the efficiency of selection. [0005] Milk production trait is an important economic trait of milk buffalo, which is determined by a variety of microgenes and is a typical quantitative trait controlled by polygenes. Clone milk buffalo lactation performance-related genes and explore their genetic characteristics to improve milk buffalo production pe...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12Q1/68
CPCC07K14/47C12Q1/6888C12Q2600/124C12Q2600/156
Inventor 邓廷贤梁贤威庞春英马小娅梁莎莎朱鹏陆杏蓉段安琴
Owner GUANGXI ZHUANG AUTONOMOUS REGION BUFFALO INST
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