Detection method and application of bull ZNF280AY (zinc finger protein 280A, Y-linked) gene copy number variations (CNVs)

A technology of gene copy number and detection method, which is applied in the field of molecular genetics, can solve problems such as undiscovered ZNF280AY gene copy number variation, and achieve the effect of convenient routine operation, accelerated breeding effect, simple and fast operation

Inactive Publication Date: 2018-12-25
LANZHOU UNIVERSITY
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Problems solved by technology

But so far, there is no report on the method for detecting the variation of the copy number of the bovine ZNF280AY gene

Method used

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  • Detection method and application of bull ZNF280AY (zinc finger protein 280A, Y-linked) gene copy number variations (CNVs)
  • Detection method and application of bull ZNF280AY (zinc finger protein 280A, Y-linked) gene copy number variations (CNVs)
  • Detection method and application of bull ZNF280AY (zinc finger protein 280A, Y-linked) gene copy number variations (CNVs)

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Embodiment 1

[0022] In this embodiment, real-time fluorescent quantitative PCR technology is used to detect the variation in the copy number of the Y chromosome ZNF280AY gene unique to bovids, which mainly includes the following steps:

[0023] 1. Collection of bull frozen semen samples and extraction of genomic DNA

[0024] 1. Collection of frozen fine tube samples

[0025] In this example, frozen semen samples of 5 breeds of Holstein, Simmental, Charolais, Limousin and Angus cattle were collected, and 3 tubes of frozen semen were collected for each sample, which were placed in liquid nitrogen during transportation. jars, and placed in a -80°C ultra-low temperature freezer in the laboratory for later use. The specific information is shown in Table 1. Among the collected frozen semen samples, 107 Simmental cattle had semen quality or testis size records, and 142 Holstein cattle had sperm quality parameters, which can be used as important indicators for the evaluation of bull reproductive...

Embodiment 2

[0100] 1. Identification of ZNF280AY gene copy number in different cattle breeds.

[0101] In the present embodiment, different cattle species (Table 3, Table 4) ZNF280AY gene copy number variation range and median copy number (Table 7, Table 4) were identified. Figure 4 ).

[0102] Table 7 The copy number variation of ZNF280AY gene in 20 cattle breeds

[0103]

[0104] Through data analysis, it was found that there were significant differences in the copy number of the ZNF280AY gene between species and within species. The median copy number in the overall test samples was 48, and the copy number varied from 11 to 154 (Table 7). Among them, Jier cattle had the highest median number of copies, 76. And the median number of copies of Anxi cattle is the lowest, which is 14 (such as Figure 4 shown). In addition, non-parametric U test was used to analyze the variation of ZNF280AY gene copy number among different Y chromosome haplotypes, and it was found that the median copy...

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Abstract

The invention provides a detection method and application of bull ZNF280AY (zinc finger protein 280A, Y-linked) gene copy number variations (CNVs). The method includes: taking bull genomic DNA (deoxyribonucleic acid) as a template and using PCR (polymerase chain reaction) to amplify primer pairs, wherein the PCR amplified primer pairs includes a primer pair P1 and a primer pair P2; respectively amplifying bull breed ZNF280 gene and SRY (sex-determining region, Y) reference gene through real-time quantitative PCR, and the copy number of the bull ZNF280AY gene is calculated based on quantitativeresults. The detection method based on fluorescence quantitative PCR has the advantages of high flexibility, low cost, simpleness and rapidless in operation and convenience in routine laboratory operation; large-sample detection can be achieved; in addition, the copy number variations and reproduction traits are subjected to correlation analysis so as to take the ZNF280AY gene as an effective molecular marker for selection of the reproduction traits of bulls, and the breeding effect of full reproduction is thus accelerated.

Description

technical field [0001] The invention belongs to the technical field of molecular genetics, and in particular relates to a detection method and application of a copy number variation of a bull ZNF280AY gene (Zinc fingerprotein 280A, Y-linked). The detection method uses qPCR technology, with the SRY gene (Sex-determining region Y) as a reference, the method of correcting the absolute quantification of the sample is used to calculate the copy number of the ZNF280AY gene in the test sample, and the correlation analysis between the copy number variation and the reproductive traits of the breeding bull . Background technique [0002] With the continuous development of molecular biology and the deepening of genome structure variation research, scholars have discovered more and more types of genome structure variation. Copy number variation (Copy Number Variations, CNVs) has become a new type of genome structure variation after microsatellites (Simple Sequence Repeat, SSR) and sing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/166
Inventor 乐祥鹏裴生伟李发弟李万宏李刚
Owner LANZHOU UNIVERSITY
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