Immunoproteomics of membrane proteins of Shigella flexneri 2a 2457T
To screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T. The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. flexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria....
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Published in | World journal of gastroenterology : WJG Vol. 11; no. 43; pp. 6880 - 6883 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Beijing Institute of Pharmaceutical Chemistry,Beijing 102205,China%Beijing Institute of Biotechnology,State Key Lab of Pathogen and Biosecurity,Beijing 100071,China
21.11.2005
Beijing Institute of Biotechnology,State Key Lab of Pathogen and Biosecurity,Beijing 100071,China Baishideng Publishing Group Inc |
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Abstract | To screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T.
The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. flexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides.
A total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine.
Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum. |
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AbstractList | R3; AIM: To screen the immunogenic membrane proteins ofShigella flexneri 2a 2457T.METHODS: The routine two-dimensional polyacrylamidegel electrophoresis (2-DE) and Western blotting werecombined to screen immunogenic proteins of S. flexneri2a 2457T. Serum was gained from rabbits immunizedwith the same bacteria. Immunogenic spots werecut out from the polyacrylamide gel and digested bytrypsin in-gel. Matrix-assisted laser desorption/ionizationtime of flight-mass spectrometry (MALDI-TOF-MS)was performed to determine the molecular weight ofpeptides. Electrospray ionization (ESI-MS/MS) wasperformed to determine the sequences of the interestingpeptides.RESULTS: A total of 20 spots were successfullyidentified from Coomassie brilliant blue stained gelsrepresenting 13 protein entries, 5 known antigens and8 novel antigens. A hypothetical protein (YaeT) wasdetected, which might be a candidate target of vaccine.CONCLUSION: Membrane proteins of S. flexneri 2a2457T were successfully observed by 2-DE. Severalknown and novel antigens were identified by massspectrum. To screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T. The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. flexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides. A total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine. Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum. AIMTo screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T.METHODSThe routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. flexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides.RESULTSA total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine.CONCLUSIONMembrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum. AIM: To screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T. METHODS: The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. flexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides. RESULTS: A total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine. CONCLUSION: Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum. |
Author | Wang, Heng-Liang Huang, Liu-Yu Huang, Pei-Tang Feng, Er-Ling Wei, Kai-Hua Ying, Tian-Yi Wang, Jun-Jun Huang, Cui-Fen |
AuthorAffiliation | Beijing Institute of Biotechnology,State Key Lab of Pathogen and Biosecurity,Beijing 100071,China;Beijing Institute of Pharmaceutical Chemistry,Beijing 102205,China%Beijing Institute of Biotechnology,State Key Lab of Pathogen and Biosecurity,Beijing 100071,China |
AuthorAffiliation_xml | – name: Beijing Institute of Biotechnology,State Key Lab of Pathogen and Biosecurity,Beijing 100071,China;Beijing Institute of Pharmaceutical Chemistry,Beijing 102205,China%Beijing Institute of Biotechnology,State Key Lab of Pathogen and Biosecurity,Beijing 100071,China |
Author_xml | – sequence: 1 givenname: Tian-Yi surname: Ying fullname: Ying, Tian-Yi organization: Beijing Institute of Biotechnology, State Key Lab of Pathogen and Biosecurity, Beijing 100071, China – sequence: 2 givenname: Jun-Jun surname: Wang fullname: Wang, Jun-Jun – sequence: 3 givenname: Heng-Liang surname: Wang fullname: Wang, Heng-Liang – sequence: 4 givenname: Er-Ling surname: Feng fullname: Feng, Er-Ling – sequence: 5 givenname: Kai-Hua surname: Wei fullname: Wei, Kai-Hua – sequence: 6 givenname: Liu-Yu surname: Huang fullname: Huang, Liu-Yu – sequence: 7 givenname: Pei-Tang surname: Huang fullname: Huang, Pei-Tang – sequence: 8 givenname: Cui-Fen surname: Huang fullname: Huang, Cui-Fen |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/16425402$$D View this record in MEDLINE/PubMed |
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Keywords | Immunoproteomics Shigella flexneri 2a 2457T Membrane proteins |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Telephone: +86-10-66948836 Correspondence to: Dr Heng-Liang Wang, Institute of Biotechnology, State Key Lab of Pathogen and Biosecurity, 20 Dongdajie Street, Fengtai District, Beijing 100071, China. wanghl@nic.bmi.ac.cn Author contributions: All authors contributed equally to the work. |
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Snippet | To screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T.
The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western... AIMTo screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T.METHODSThe routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and... R3; AIM: To screen the immunogenic membrane proteins ofShigella flexneri 2a 2457T.METHODS: The routine two-dimensional polyacrylamidegel electrophoresis (2-DE)... AIM: To screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T. METHODS: The routine two-dimensional polyacrylamide gel electrophoresis (2-DE)... |
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SubjectTerms | Animals Antigens, Bacterial - chemistry Antigens, Bacterial - immunology Bacterial Proteins - chemistry Bacterial Proteins - immunology Child Humans Membrane Proteins - chemistry Membrane Proteins - immunology Proteomics - methods Rabbits Rapid Communication Shigella flexneri - immunology |
Title | Immunoproteomics of membrane proteins of Shigella flexneri 2a 2457T |
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