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A Molecular Marker Method for Selecting Goat Litter Size Using a Delinked Metalloprotease Gene Containing a Thrombospondin Motif

A metalloprotease and sensitive protein technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problem of female reproductive capacity reduction, achieve the effect of accelerating breeding progress and improving selection efficiency

Active Publication Date: 2020-04-07
河南省动物卫生监督所
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Problems solved by technology

In conclusion, ADAMTS1 deficiency results in reduced female fertility but does not affect female estrous cycle and implantation of fertilized eggs

Method used

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  • A Molecular Marker Method for Selecting Goat Litter Size Using a Delinked Metalloprotease Gene Containing a Thrombospondin Motif
  • A Molecular Marker Method for Selecting Goat Litter Size Using a Delinked Metalloprotease Gene Containing a Thrombospondin Motif
  • A Molecular Marker Method for Selecting Goat Litter Size Using a Delinked Metalloprotease Gene Containing a Thrombospondin Motif

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Experimental program
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Effect test

Embodiment Construction

[0025] A. PCR amplification of exon 2 of ADAMTS1 gene and detection of its polymorphism

[0026] 1. Collection and processing of goat blood samples

[0027] Take 5 mL of goat blood sample, add 0.2 μL of ACD (2.4 g of citric acid, 6.6 g of trisodium citrate, 7.35 g of glucose, dilute to 50 mL, autoclave) for anticoagulation, slowly invert 3 times and put it in an ice box, - Store at 80°C for later use.

[0028] In this example, a total of 506 ewe blood samples from 2 goat populations were used, specifically: 263 blood samples from Xinong Saanen sheep; 243 blood samples from Henan dairy goats, collected from dairy goat breeding base in Henan Province.

[0029] 2. Extraction and purification of genomic DNA from blood samples

[0030] (1) Thaw the frozen blood sample at room temperature, transfer 500μL to a 1.5mL Eppendorf tube, add an equal volume of PBS buffer, mix well, centrifuge at 12000r / min for 10min (4℃), discard the supernatant, repeat the above steps to the supernatant...

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Abstract

The invention discloses a molecular marking method for selecting the goat kidding number by using a de-linked metalloprotease gene containing a thrombin-sensitive protein die body. The method adopts the DNA sequence of a goat genome as a template. With the presence of a taq DNA polymerase, a buffer environment, Mg2+ and dNTPs, the gene exon 2 of a de-linked metalloprotease (ADAMTS 1) containing the thrombin-sensitive protein die body is amplified with the presence of a primer P1 in the PCR condition. Based on the DNA sequencing technique, the single nucleotide polymorphism (SNP) of the exon 2 is detected. Gene sequencing results show that, the amplification sites of the primer P1 contains one base mutation, and then the amplified product of the primer P1 is subjected to genetic typing and gene frequency analysis. Meanwhile, the association analysis is conducted on the kidding traits of Henan dairy goats and the kidding traits of Xinong Saanen dairy goats. Results show that, the SNP sites of the gene exon 2 of the de-linked metalloprotease (ADAMTS 1), detected by the primer P1, can be used as molecular markers for selecting the kidding number traits of goats.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and in particular relates to a molecular marker method for selecting goat litter size by using a de-separated metalloprotease gene containing a thrombin-sensitive protein motif. Background technique [0002] With the rapid development of molecular biology technology, especially the advent of Polymerase Chain Reaction (Polymerase Chain Reaction) technology and the improvement of electrophoresis technology, various molecular genetic marker technologies have emerged. The cultivation of provides new ways and methods. In particular, the combination of molecular marker technology and conventional breeding technology has given birth to a brand-new selection method, that is, marker-assisted selection (Marker assistant selection). It can overcome the environmental deviation and sampling error caused by phenotypic selection in the traditional selection process, improve the accuracy and efficiency of live...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 白跃宇谭旭信白怡霖马云牛桂玲
Owner 河南省动物卫生监督所
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