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Gene srebp1 related to buffalo lactation and its application as a molecular marker

A buffalo and genetic technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problem of no SREBP1 gene, etc., achieve high milk protein rate and improve production performance

Active Publication Date: 2020-06-12
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 SREBP1 gene as a molecular marker for buffalo milk production traits

Method used

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  • Gene srebp1 related to buffalo lactation and its application as a molecular marker
  • Gene srebp1 related to buffalo lactation and its application as a molecular marker
  • Gene srebp1 related to buffalo lactation and its application as a molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: Preparation of buffalo genome total DNA

[0031] 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:

[0032] (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 procedure Step once; add 4μL RNAase (100mg / mL), shake for 15sec, and let stand at room temperature for 5min;

[0033] (2) Add 20 μL of Proteinase K solution, mix well, add 200 μL of buffer GB, fully invert and mix, place at 56°C for 10 min, invert and mix several times during the period, the solution should become clear;

[0034] (3) Add 200 μL of absolute ethanol and mix thoroughly by inversion; flocculent precipitation may appear at this time;

[0035] (4) Add the solution obtain...

Embodiment 2

[0041] Example 2: Acquisition of SREBP1 Gene Molecular Marker for Buffalo Milk Production Traits

[0042] 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 SREBP1 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.

[0043] 2. PCR amplification reaction

[0044] Genomic DNA samples from 50 buffaloes were randomly selected for pooling as templates for PCR reactions.

[0045](1) PCR reaction: The total reaction volume is 40 μL, including 1 μL of buffalo DNA pool template, 20 μL of 10X LA Taq Mix, 2 μL of primer mixture (10 mM each) and ddH 2 O 17 μL.

[0046] (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

[0047] 3. PCR product sequencing, analysis and molecular ...

Embodiment 3

[0063] Example 3: Gene SREBP1 related to buffalo lactation and its application as a molecular marker

[0064] The breed of the test buffalo herd was a hybrid milk buffalo herd. The association analysis between different genotypes of SREBP1 gene C8358T and G15306T loci and milk production traits was carried out in buffalo herds.

[0065] The results of association analysis of buffalo SREBP1C8358T and G15306T loci 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). For the C8358T locus, the milk yield of buffalo individuals carrying CC genotype was higher than that of TC and TT individuals. For the G15306T locus, the milk production of buffalo individuals carrying the GT genotype was higher than that of individuals with other genotypes. For G15306T, the milk protein rate of buffalo individuals carrying GT genotype was higher than that of individuals with other genotypes. Fo...

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Abstract

The invention provides a SREBP1 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 C8358T and G15306T single-base mutation occurs at site 8358 bp and site 15306 bp, respectively; the SNP site at the 8358 bp is substantially related to the milk output of the buffalo; and the SNP site at the 15306 bp is substantially related to the milk output, milk protein rate and lactose rate 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 SREBP1 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 high milk output, high milk protein rate and high lactose rate of produced milk.

Description

【Technical field】 [0001] The invention relates to the technical field, in particular to the field of animal molecular markers. It specifically relates to the gene SREBP1 related to buffalo lactation and its application as a molecular marker. 【Background technique】 [0002] 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. [0003] 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 performance, pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12Q1/6888
CPCC07K14/47C12Q1/6888C12Q2600/124C12Q2600/156
Inventor 邓廷贤梁贤威马小娅庞春英梁莎莎陆杏蓉朱鹏段安琴
Owner GUANGXI ZHUANG AUTONOMOUS REGION BUFFALO INST
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