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Method for labeling escherichia coli proteome by using SILAC (Stable Isotope Labeling with Amino Acids in Cell Cultures) and special culture medium
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A technology of Escherichia coli and culture medium, applied in the fields of peptide preparation methods, microorganism-based methods, chemical instruments and methods, etc., can solve the problems of hindering the progress of Escherichia coli quantitative proteomics, lack of labeling methods, etc., and achieve the elimination of genetic conditions. The effect of limiting, eliminating tedious steps, and efficient marking
Active Publication Date: 2013-09-04
ACADEMY OF MILITARY MEDICAL SCI
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Due to the lack of effective labeling methods, there has not been the advent of E. coli high-efficiency SILAC medium and the publication of large-scale quantitative proteomics research papers, which hinders the progress of E. coli quantitative proteomics research and its application in the fermentation industry and basic medical research. play a role in
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Embodiment 1
[0049] Embodiment 1, SILAC marker Escherichia coli special culture medium
[0050] Table 1 is the composition of the special medium for SILAC-labeled Escherichia coli
[0051]
[0052] Remarks: Inorganic reagents are ordinary analytical pure domestic reagents, light-labeled amino acids were purchased from Amresco Company in the United States, L-lysine- 13 C 6 Purchased from American CIL Company, article number: CNLM-291-0.25.
[0053] According to the above-mentioned components and their final concentrations, prepare with ultrapure water to obtain a special medium for SILAC-labeled Escherichia coli. Inorganic salts and glucose are sterilized by autoclaving, and amino acids and vitamins are sterilized by filtration.
Embodiment 2
[0054] Embodiment 2, the application of SILAC marker Escherichia coli special medium
[0055] 1. SILAC-labeled E. coli special medium to mark E. coli
[0056] 1. SILAC mark
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Abstract
The invention discloses a method for labeling escherichia coli proteome by using SILAC (Stable Isotope Labeling with Amino Acids in Cell Cultures) and a special culture medium. The invention provides a special culture medium for the SILAC-labeled escherichia coli. The special culture medium comprises inorganic salt, histidine hydrochloride, isoleucine, valine, leucine, phenylalanine, tryptophan, serine, heavy stable isotope labeled lysine, arginine, threonine, tyrosine, methionine, adenine sulfate, uracil, vitamin B1 and glucose; and the inorganic salt is Na2HPO4.12H2O, KH2PO4, NaCl, MgSO4, NH4Cl and CaCl2. Experiments prove that, the special culture medium for the SILAC-labeled escherichia coli, disclosed by the invention, creates a condition for researches of labeling the escherichia coli proteome, preparing a quantitative internal label and highly-precisely quantifying the escherichia coli proteome.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a method for using SILAC to mark Escherichia coli proteome and a special culture medium thereof. Background technique [0002] Escherichia coli is the abbreviation of Escherichia coli (Escherichia coli), named after it was discovered by Theodor Escherich in 1885. Escherichia coli Bacteria, Proteobacteria, Gammaproteobacteria, Enterobacteriales, Enterobacteriaceae, Escherichia Escherichia coli is a Gram-negative short bacillus with a size of 0.5×1-3 microns, with flagella all over the body, able to move, and without spores. [0003] Escherichia coli is widely distributed in nature, and it is also the most important and most abundant bacterium in the intestines of humans and many animals. It mainly parasitizes in the large intestine, most of which do not cause disease, and accompany people for life. Its metabolic activity can inhibit the growth of microorganisms that decompose protei...
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