Sample preparation and treatment method for researching metaproteomics of soil

Abstract

The invention discloses a sample preparation and treatment method for researching the metaproteomics of soil. The method comprises the following steps: pretreating a soil sample to obtain a first suspension; carrying out concussion crushing to obtain a crushed first suspension; carrying out ultrasonic treatment, centrifuging the treated suspension, and collecting the obtained first supernatant; precipitating the first supernatant with cold acetone to obtain a protein solution; precisely weighing the protein solution containing proteins, adding a single protease or a protease combination, and incubating the proteins overnight to obtain a polypeptide solution; adding a separating agent 1, carrying out vortex treatment, adding a separating agent 2, carrying out vortex treatment, centrifuging the obtained solution, standing the centrifuged solution, and transferring about 90% of the obtained lower layer phase of a polypeptide-containing segment into a new centrifuging tube; adding a separating agent 3, carrying out vortex treatment to realize uniform mixing, centrifuging the treated solution, and taking the obtained lower layer; mixing the above obtained two peptide segment solution, desalting the obtained solution mixture, and collecting the obtained eluate; and centrifuging, concentrating and drying the eluate, afresh dissolving the obtained peptide segment sample, and directly injecting the dissolved sample to a machine for analysis. The method has the advantages of simplicity in operation, high stability, high recovery rate, completeness in extraction, and high flux, and is of great significance to massively develop the relevant researches of the metaproteomics of soil.

Description

technical field [0001] The invention relates to a method for extracting and processing whole proteins for the purpose of proteomics analysis, especially a method for soil [0002] Sample preparation and processing methods for soil macroproteomics research. Background technique [0003] With the increasing maturity and development of soil metagenomics, soil metaproteomics, as an important technology platform in the post-genome era, has attracted more and more attention. Soil protein plays an important role in the biogeochemical cycle of carbon, nitrogen and phosphorus and the accumulation of organic matter, and is closely related to soil quality and sustainable development. Proteomics essentially refers to the study of protein characteristics on a large scale, including protein expression levels, post-translational modifications, and protein-protein interactions. Metaproteomics is a technical means to study the variation of protein species and abundance from all microbial s...

AI Technical Summary

Problems solved by technology

The main reasons are as follows: 1. The matrix of the soil is very complex, which interferes with protein extraction; 2. There are many kinds of proteins with large differences in physical properties, and the current methods are difficult to achieve efficient extraction of all proteins; 3. The abundance of protein in the soil There is a large difference in concentration, some protein content is very small, and the existing methods often miss low-abundance proteins; 4. The current processing methods are generally cumbersome, and pollution and uncertain factors are likely to be caused during the process; 5. With the As the number of samples to be analyzed increases, the throughput of the processing method is also particularly important

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