Cloning of buffalo milk production character MC4R gene and application thereof serving as molecular marker

A molecular marker and buffalo technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problem that there is no molecular marker of buffalo MC4R gene, etc., and achieve the effect of high-throughput detection of samples

Active Publication Date: 2014-07-16
GUANGXI ZHUANG AUTONOMOUS REGION BUFFALO INST
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there are no molecular markers for

Method used

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  • Cloning of buffalo milk production character MC4R gene and application thereof serving as molecular marker
  • Cloning of buffalo milk production character MC4R gene and application thereof serving as molecular marker
  • Cloning of buffalo milk production character MC4R gene and application thereof serving as molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Acquisition of gene MC4R related to milk production traits in buffalo

[0036] 1. Preparation of total genome DNA of buffalo

[0037] Genomic total DNA of buffalo was prepared by blood genomic DNA extraction kit (Tiangen, Beijing),

[0038] The specific operation steps are as follows:

[0039] (1) Take 500 μL of whole blood and place it in a 1.5mL centrifuge tube, add an equal volume of cell lysate CL, mix by inversion, centrifuge at 12,000rpm 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;

[0040] (2) Add 20 μL Proteinase K solution, mix well, add 200 μL buffer GB, fully invert and mix, place at 56°C for 10 minutes, invert and mix several times during the period, the solution should become clear;

[0041] (3) Add 200 μL of absolute ethanol, and mix thoroughly by inversion; flocculent precipitatio...

Embodiment 2

[0067] Screening and Detection of Molecular Markers of Buffalo MC4R Gene

[0068] 1. Preliminary screening of buffalo MC4R gene molecular markers

[0069] According to the buffalo family pedigree and milk production records, select 5 high-yielding buffaloes (≥2000kg / lactating period) and 5 low-yielding buffaloes (≤1000kg / lactating period) as the primary screening objects of molecular markers; using SEQ ID NO: 2-7 The primer sequences of the above 10 buffalo DNA samples were amplified by PCR, and 5 μL of PCR amplification products were taken for agarose gel electrophoresis detection, and the remaining samples were sent to Shenzhen Huada Technology Co., Ltd. for sequencing; DNAStar7.0Seqman bioinformatics analysis software was used to analyze The sequencing results were spliced ​​and compared, and the molecular markers of the MC4R gene related to buffalo milk production traits were initially screened (see Table 1). There were 13 base mutations in total, 5 of which were located i...

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Abstract

The invention discloses cloning of a buffalo milk production character MC4R gene and an application thereof serving as a molecular marker. A sequence obtained by cloning is as shown in SEQ ID NO:1, three base mutations affecting the buffalo milk production character are detected, which are respectively C226-T226 at 226bp, C1104-T1104 at 1104bp and A1525-G1525 at 1525bp. The invention discloses a buffalo gene MC4R sequence, 3 base mutation sites affecting the buffalo milk production character, a method for obtaining the gene and an application thereof, and provides a new molecular marker for auxiliary selection of buffalo milk production character markers.

Description

technical field [0001] The invention belongs to the technical field of animal genetic engineering, and in particular relates to the cloning of the MC4R gene of buffalo milk production traits 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, provide techn...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/12C12N15/10
CPCC07K14/72C12Q1/6888C12Q2600/124C12Q2600/156
Inventor 庞春英邓廷贤梁贤威杨炳壮刘满清陈明棠黄健谭正准李辉
Owner GUANGXI ZHUANG AUTONOMOUS REGION BUFFALO INST
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