Method for efficiently enriching cell protein of buffalo

Abstract

The invention relates to the technical field of protein extraction, in particular to a method for efficiently enriching cell protein of a buffalo. The method comprises the steps of cell pyrolysis, preparation of a protein sample, purification of protein, recycling of peptide fragments and the like, the method of combining non-contact ultrasonic sample crushing and protein enrichment by magnetic beads is adopted, the sample dosage is greatly lowered, and the extracting efficiency of protein is improved. The method has high operating practicability and is easy to popularize, an effective sampletreatment method is provided for a proteomic technology to study micro-samples or precious samples so that the method can satisfy the sample condition needed by the proteomic study. By means of the sample treatment method, the bottleneck problem that currently, the micro-samples cannot satisfy the sample dosage needed by the proteomic study is solved, a feasible scheme is also provided for the unicellular proteomic analysis, and the cell protein of the buffalo is efficiently enriched.

Description

【Technical Field】 [0001] The invention relates to the technical field of protein extraction, in particular to a method for efficiently enriching buffalo cell protein. 【Background technique】 [0002] Chinese buffaloes are all swamp-types, which were originally draught animals in the southern region. Later, with the mechanization of agriculture, the value of buffalo milk gradually replaced its draught value. However, the low reproductive efficiency of Chinese buffalo made its milk value less Development, and the quality of germ cell maturation is one of the main factors affecting the reproductive rate of buffalo. This complexity is mainly manifested in transcription, translation, post-translational modification and regulatory networks. Proteins are the executors of life functions. The use of high-throughput proteomics technology can discover protein expression and post-translational modifications on a large scale, which helps to reveal the protein regulatory network of germ cell de...

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Problems solved by technology

[0003] In 2014, Daniela R. Deutsch et al identified 1072 proteins using 1600 oocytes or early embryos; in 2015, Chen Fumei et al identified 574 proteins using 1000 buffalo oocytes in GV and MII stages; In 2016, Chen Lingsheng and others identified 647 and 570 proteins from 522 GV stage and 545 MII stage buffalo oocytes respectively. Buffalo oocytes and early embryo materials are extremely precious. The amount of sample required requires thousands of cells, and the conventional protein separation and purification methods cannot efficiently enrich the trace protein of buffalo oocytes and early embryos, which brings certain difficulties to the research of buffalo reproduction. Therefore, it is urgent to find out a A method that can efficiently enrich trace proteins in buffalo oocytes and early embryos can effectively improve the efficiency of protein extraction and meet the requirements of the sample volume required for analysis, thereby realizing the proteomics research of buffalo trace biological samples and even single cell proteins

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