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A method for mining differential proteins in testes of Tibetan sheep before and after sexual maturity based on dia technology

A digging and hidden sheep technology, applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of poor reproducibility of peptide identification

Active Publication Date: 2022-04-01
GANSU AGRI UNIV
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AI Technical Summary

Problems solved by technology

In the conventional data-dependent acquisition mode, the randomness of precursor ion acquisition will cause poor reproducibility of peptide identification

Method used

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Embodiment 1

[0030] A method for mining differential proteins in Tibetan sheep testes before and after sexual maturity based on DIA technology, comprising the following steps:

[0031] Step A. Tissue collection

[0032] In the described step A, the testicular tissues described in this patent come from male Tibetans with similar genetic background (same paternal mother) at two key developmental stages before sexual maturity (3 months old) and after sexual maturity (1 year old). Sheep: After slaughtering the sheep, take out the testis tissue, wash the external blood stains with PBS and peel off the buffy coat, cut it into small tissues of 0.5cm×0.5cm×0.5cm, and quickly freeze them in liquid nitrogen for protein samples preparation of

[0033] Step B. Protein sample preparation

[0034] Add lysate (7M urea, 2% SDS, 0.1% PMSF, 65mMDTT) to the testicular tissue, and ultrasonically lyse; after centrifuging at 4°C and 14000rpm for 30min, draw the supernatant, and use the BCA protein quantificat...

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Abstract

The invention discloses a method for mining differential proteins in testis of Tibetan sheep before and after sexual maturity based on DIA technology, comprising the following steps: tissue collection, protein sample preparation, spectral database establishment, protein qualitative and quantitative analysis, and differentially expressed protein screening. The present invention selects the testicular tissues of 3-month-old and 1-year-old Tibetan sheep, uses conventional DDA mass spectrometry detection technology to analyze and establish a map database, and then adopts DIA method to collect mass spectrometry data, and conducts qualitative and quantitative analysis on the protein in the sample, so as to obtain Tibetan Differentially expressed proteins in sheep testis. Using this method, the quantitative screening of differentially expressed proteins in Tibetan sheep testes can be achieved, which provides a very important theoretical basis for the subsequent study of the molecular mechanism of Tibetan sheep testis development or spermatogenesis from the perspective of proteomics.

Description

technical field [0001] The invention relates to the technical field of DIA quantitative methods, in particular to a method for mining differential proteins in the testes of Tibetan sheep before and after sexual maturity based on the DIA technology. Background technique [0002] As one of the three primitive sheep breeds in China, Tibetan sheep are mainly distributed in Tibet, Qinghai, Gansu, Sichuan and other places on the Qinghai-Tibet Plateau above 3000m above sea level. Sustainable development and maintaining ecological balance play a very important role. However, due to long-term grazing and lack of supplementary feeding, their production performance is low, their fecundity is low, and their sexual maturity is late, which seriously limits the reproduction of Tibetan sheep flocks, and thus restricts the economic and social development of pastoral areas. Proteins are the main participants and executors in the life activities of organisms. Therefore, it is of great signif...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 李讨讨马友记王霞张宏豫
Owner GANSU AGRI UNIV
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