Rapid extraction method for detection of outer membrane protein F of Pseudomonas aeruginosa |
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| Affiliation: | 1. Department of Veterinary Preventive Medicine and Animal Reproduction, São Paulo State University (Unesp), School of Agricultural and Veterinarian Sciences, Jaboticabal, Brazil;2. Department of Veterinary Pathology, São Paulo State University (Unesp), School of Agricultural and Veterinarian Sciences, Jaboticabal, Brazil;3. University of Guelph, Department of Pathobiology, 50 Stone Rd. East, Guelph, Ontario, N1G 2W1, Canada;4. Animal Health Laboratory, University of Guelph, Post Office 3612, Guelph, Ontario, N1H 6R8, Canada;5. Brazilian Agricultural Research Corporation (EMBRAPA), Embrapa Southeast Livestock, Washington Luiz road, Km 234, 13560-970, Sao Carlos, Sao Paulo, Brazil;1. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China;2. National Institutes for Food and Drug Control, Beijing 100050, China;1. Ege University, Department of Analytical Chemistry, Faculty of Pharmacy, 35100, Bornova, Izmir, Turkey;2. Yuzuncu Yil University, Department of Analytical Chemistry, Faculty of Pharmacy, 65010, Van, Turkey;1. Department of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, Canada;2. Department of Physiology, Faculty of Medicine, University of Alexandria, Alexandria, Egypt;3. Department of Chemistry and Biochemistry, University of Regina, Regina, SK, Canada;3. Departments of Molecular Microbiology, Cuernavaca, Morelos 62210, México;4. Molecular Medicine and Bioprocesses, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62210, México |
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| Abstract: | A filtration technique has been developed to trap the peptidoglycan sacculus for rapid extraction of the outer membrane protein F of Pseudomonas aeruginosa. The method consists of the following three steps: (i) removal of cell components except peptidoglycan associated with some proteins, (ii) trapping of peptidoglycan with a glass filter and (iii) extraction with a hot SDS solution of proteins, mainly F protein, associated with peptidoglycan. The method is simple and rapid, providing for efficient screening for protein F-deficient mutants of P. aeruginosa. Using this method of screening, three mutants were isolated among 500 mutagenized clones. |
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