Extraction method of porcine streptococcus total protein in biofilm state

Abstract

The invention relates to an extraction method of a porcine streptococcus total protein in a biofilm state, which comprises the following steps: (1) treating porcine streptococcus in a biofilm state; (2) heavily suspending the cleaned thallus cell obtained in the step (1) with a sucrose buffer into which mutanolysin is added, and incubating at 36.5-37.5 DEG C; (3) centrifuging for 8-15 minutes while collecting protoplastic cells, carrying out ultrasonic treatment on the heavily suspended cells in the buffer, adding a proteinase inhibitor, and carrying out ultrasonic crushing for 60-100 cycles in an ice bath; (4) incubating at 15-32 DEG C for 10-60 minutes; (5) adding 5-15% TCA into the protein supernatant, and applying an ice bath; and washing the protein with precooled acetone, centrifuging to collect the total protein, and drying the collected total protein at room temperature, thereby obtaining the bacterium total protein. The method provided by the invention can be widely used for extracting various bacterium total proteins in the biofilm state, and especially has wide application prospects in researching bacterium protein groups in the bacterium biofilm state.

Description

technical field [0001] The invention relates to a method for extracting bacterial total protein in the biofilm state in the field of proteomics research. Background technique [0002] With the completion of the genome sequencing of many species, people's attention began to turn to how to explain the information in the genome sequence from the perspective of structure, function and control of biological systems. The emergence of proteomics and bioinformatics has opened up new ways to study complex biological systems. The most widely used technology in proteomics research and the most reliable technology in parallel experiments is two-dimensional electrophoresis technology, followed by biological mass spectrometry identification of separated proteins (Gorg A, Weiss W, Dunn MJ. Current two-dimensional electrophoresis technology for proteomics . Proteomics, 2004, 4: 3665-3685.). Since various protein extraction methods used for two-dimensional electrophoresis often vary greatl...

AI Technical Summary

Problems solved by technology

In the phenol extraction method, taking advantage of the property that protein is soluble in Tris saturated phenol, the impurities insoluble in organic solvents can be effectively removed by adding the extract and repeated extraction, and most of the salt ions are left in the extraction process. Extraction solution, thus playing a certain degree of salt removal (Li XF, Han HP, Wang XC, Fan PX and Li YX. Extraction methods for two-dimensional electrophoresis analysis of shoot proteins in halophyte Salicornia europaea. Acta Ecologica Sinica, 2006 , 26: 1848-1853.), but in cells, polyphenols will irreversibly combine with proteins through hydrogen bonds, resulting in flaky dispersion in the two-dimensional electrophoresis gel image, and some insoluble carbon compounds will also block IPG gel The gel wells of the strip, thus prolonging the focusing time, eventually leading to tailing and partial protein loss

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