Method for extracting mycoplasma bovis biofilm total protein

A technology of mycoplasma bovis and biofilm, applied in the field of protein extraction, to achieve the effect of overcoming incomplete separation, clear image and sufficient aggregation

Inactive Publication Date: 2016-11-30
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the prior art, there is no effective extraction method for Mycoplasma bovis biofilm

Method used

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  • Method for extracting mycoplasma bovis biofilm total protein

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Embodiment Construction

[0015] Below in conjunction with embodiment for explaining in detail the extraction method of mycoplasma bovis biofilm total protein of the present invention, involved in implementing and test instrument are:

[0016] The first isoelectric focusing instrument Ettan IPGphor Isoelectric Focusing System, the second vertical plate electrophoresis instrument Hofer SE 600, and the power supply EPS-601 are all products of GE Amersham; the pure water device is a product of Millpore; the image scanner Typhoon FLA9000 is a product of GE product. IPG dry strips (13cm, pH3-10), IPG Buffer (pH3-10), CHAPS, dithiothreitol (DTT), and iodoacetamide were all purchased from GE Healthcare in the United States; SDS, Glycine, Tris, Acrymide, BIS, APS, Urea, and TEMED are all BIO-RAD reagents; protein quantification kits are products of Beijing Regen Biotechnology Co., Ltd.; absolute ethanol, acetone, and trichloroacetic acid are all domestic analytical grade; other reagents are all products of GE ...

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Abstract

The invention discloses a method for extracting mycoplasma bovis biofilm total protein capable of shortcomings in the prior art. The method for the extracting mycoplasma bovis biofilm total protein comprises the steps of first adopting a mechanical method to collect mycoplasma bovis biofilm thalli and then adding lysate into the mycoplasma bovis biofilm thalli to perform pyrolysis, causing mycoplasma bovis having undergone pyrolysis to undergo ice-bath ultrasonic treatment and then separating supernatant, finally adding TCA / acetone solution into the supernatant obtained in the last step, and after low-temperature precipitation, removing the supernatant to obtain the total protein. According to the method, the problems that in the prior art, polysaccharide ingredients and the thalli are separated incompletely, polysaccharides and DNAs and the like remain and the total protein degrades and loses easily are solved; the step of removing impurities such as lipid matter in the sample preparation process (removing the lipid matter through the TCA / acetone solution) is added, the interference of nonprotein matter is reduced in the subsequent process of isoelectric focusing electrophoresis of the extracted total protein sample is reduced, and the total protein is effectively separated in the isoelectric focusing and polyacrylamide gel electrophoresis process.

Description

technical field [0001] The invention relates to a method for extracting protein, specifically, the invention relates to a method for extracting total protein of mycoplasma bovis biofilm. Background technique [0002] At present, life science research has entered the post-genome era, and proteomics has become the frontier and research hotspot of studying organisms in the post-genome era. Two-dimensional electrophoresis is the most classic proteomics technique. Through two-dimensional electrophoresis, protein samples (including microbial cells, cells, tissues, etc.) are separated according to the isoelectric point and molecular weight, and then the separated protein is identified by mass spectrometry. qualitative and quantitative information about a protein. In two-dimensional electrophoresis technology, the extraction and preparation of proteins is very important. The quality of protein extraction and preparation directly affects the effect of two-dimensional electrophoresi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/30
Inventor 陈胜利郝华芳储岳峰赵萍刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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