A method for efficiently enriching protein in buffalo cells
A protein and water buffalo technology, applied in the field of protein extraction, can solve problems such as the difficulty of buffalo reproduction research, the inability of protein separation and purification methods to efficiently enrich trace proteins, etc., and achieve the effects of rapid magnetic responsiveness and high carboxyl content.
Active Publication Date: 2022-04-05
GUANGXI UNIV
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[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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[0050] The method for purifying buffalo cell protein in this embodiment is as follows: first pretreat the magnetic beads to obtain a homogeneous magnetic bead solution, then enrich the buffalo cells and put them on ice for the first lysis; Add a mixture of homogeneous magnetic bead solution and trifluoroethanol to buffalo cells and perform a second lysis; then perform protein purification and peptide recovery on the second lysate to obtain enriched buffalo cell proteins;
[0051] The above-mentioned buffalo cells are buffalo oocytes or buffalo early embryo cells; wherein, the buffalo early embryo cells include parthenogenetic activated embryo cells and in vitro fertilized embryo cells:
[0052] The buffalo ovary used in this embodiment is provided by Nanning Roulian Food Processing Factory;
[0053] 1. Cell collection:
[0054] After the buffalo ovary is sent to the laboratory, remove excess tissue with scissors, and wash it with double-antibody saline at about 39°C for 2-3 t...
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The present invention relates to the technical field of protein extraction, in particular to a method for efficiently enriching buffalo cell proteins. The method includes the steps of cell lysis, protein sample preparation, protein purification and peptide segment recovery, and uses non-contact ultrasonic crushing of samples combined with magnetic beads. The method of protein enrichment greatly reduces the amount of sample used and improves the extraction efficiency of protein. This technology is practical and easy to popularize. The sample processing method of the invention can meet the sample conditions required for proteomics research. The sample processing method of the invention solves the bottleneck problem that the current micro-sample cannot meet the sample size required for proteomics research. The analysis provides a feasible solution and is an efficient method for enriching buffalo cell proteins.
Description
【Technical field】 [0001] The invention relates to the technical field of protein extraction, in particular to a method for efficiently enriching buffalo cell proteins. 【Background technique】 [0002] Chinese buffaloes are all swamp type, and they were originally draft animals in the southern region. Later, with the mechanization of agriculture, the value of milk and meat of buffaloes gradually replaced their value for labor. However, the low reproductive efficiency of Chinese buffaloes also prevented the value of milk and meat from being well-received. Development, while the maturation quality of germ cells is one of the main reasons affecting the reproductive rate of buffalo, this complexity is mainly manifested in transcription, translation and post-translational modification and regulatory network. Proteins are the executors of life functions. High-throughput proteomics technology can be used to discover protein expression and post-translational modifications on a large s...
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IPC IPC(8): C07K1/22C07K1/14C07K1/36
CPCC07K1/14C07K1/22C07K1/36
Inventor 张明濮黎萍陈富美
Owner GUANGXI UNIV