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Detection of Bacteria and Bacteriophage Proteins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Abstract
Rapid and reliable identification of microorganisms is of paramount importance for advancing homeland security. Mass spectrometry is emerging as an effective bioanalytical tool having unique capabilities in handing complex mixtures. Matrix-assisted layser desorption / ionization time-of-flight mass spectrometry ¡]MALDI/TOF MS¡^is commonly used in the analysis of bio-molecules owing to its merits of high sensitivity, wide mass range, and ease-of-operation.
The objective of the first part of the thesis is to develop an analysis method to characterize the genus, strains, and subspecies of the infections gastroenteritis bacteria by using MALDI/TOF MS. MALDI/TOF MS can be applied to classify different microoganisms based on examination of ion patterns from different microoganisms and unique distinguishing protein ions as biomarkers for different strains. The second section, bacteriophages MS2 and T4 were used as materials for identification of their specific protein markers. Protocols including sample preparation, purification, combined in gel digestion and post source decay to which the mass spectrometer brought for analytical specificity were employed.
According to the experimental results, classification of different microorganisms based on examination of ion patterns is feasible. Furthermore, the peak at m/z 5255 in the MALDI mass spectra of MS2 analysis and the peak at m/z 8600 of T4 can provide the unique distinguishing signals. The experimental mass spectral peaks were submitted to the database search, and one of these peaks was matched to a tail fiber assembly helper protein. Thus, the method developed and the mass spectra presented in this thesis can be potentially applied to the practical virus analysis.

Identiferoai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0902104-153708
Date02 September 2004
CreatorsLee, Jen-Yi
ContributorsJentaie Shiea, Yi-Yang Lien, Ming-Hong Tai, Chung-Lung Cho
PublisherNSYSU
Source SetsNSYSU Electronic Thesis and Dissertation Archive
LanguageCholon
Detected LanguageEnglish
Typetext
Formatapplication/pdf
Sourcehttp://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0902104-153708
Rightsoff_campus_withheld, Copyright information available at source archive

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