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Proteome Analysis of Bacillus Subtilis by Two-Dimensional Difference Gel Electrophoresis and Mass Spectrometry

Author: JiaFengFeng
Tutor: ZhaoXueMing
School: Tianjin University
Course: Biochemical Engineering
Keywords: Proteomics Bacillus subtilis Two - dimensional gel electrophoresis Riboflavin Key enzyme
CLC: Q936
Type: Master's thesis
Year: 2009
Downloads: 70
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Abstract


Proteomics is the study of genome expression of all proteins in a discipline , is an important part in the post-genomic era group studies , have achieved very important results in different cross-cutting areas . Bacillus subtilis as an important model organism of Gram- positive bacteria , not only in the field of basic biology with important applications , is also a very important industrial microorganisms . Therefore, this article attempts to proteomics approach differences in research on genetically engineered Bacillus subtilis and the original strain . Proteomics research for the experimental use classic method ( two-dimensional gel electrophoresis ) of the two strains proteome analysis . The specific use of experimental techniques : sample preparation techniques ; two-dimensional gel electrophoresis separation technology ; gel staining techniques ; image analysis techniques ; identified by mass spectrometry technology and bioinformatics technology . The experimental two bacterial protein expression profile differences proteome analysis and focus on the metabolic pathways of riboflavin production , some important metabolic pathways related information used to spawn the next step transformation . The specific results : ( 1 ) the use of gel electrophoresis separation method identified a total of 840 the protein points , respectively encoded by 542 genes . 83 proteins which appear more than twice the cut , more than twice the 75 proteins raised , the rest of the protein changes were twice within . ( 2 ) the experimental from proteomics angle validation Bacillus subtilis riboflavin metabolism transformation results , specific data related protein expression . (3) The emphasis in the analysis of this experiment riboflavin metabolism related several ways , starting from glycolysis until riboflavin synthesis, derived direction of Next strains transformation by the combination of the expression of key enzymes and metabolic pathways , should be to improve bacterial synthesis of RNA, protein , and the ability to utilize carbon sources , and in a few specific sites on the transformation to strengthen the pathway of riboflavin generate several restriction sites , and ultimately contribute to the synthesis of riboflavin .

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CLC: > Biological Sciences > Microbiology > Microbial biochemistry
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