Detailed Information on Publication Record
2011
Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin
BENEŠÍK, Martin, Jiří NOVÁČEK, Lubomír JANDA, Radka DOPITOVÁ, Lukáš ŽÍDEK et. al.Basic information
Original name
Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin
Authors
BENEŠÍK, Martin (203 Czech Republic, belonging to the institution), Jiří NOVÁČEK (203 Czech Republic, belonging to the institution), Lubomír JANDA (203 Czech Republic, guarantor, belonging to the institution), Radka DOPITOVÁ (203 Czech Republic, belonging to the institution), Lukáš ŽÍDEK (203 Czech Republic, belonging to the institution), Jiří DOŠKAŘ (203 Czech Republic, belonging to the institution), Vladislava RŮŽIČKOVÁ (203 Czech Republic, belonging to the institution), Marek MOŠA (203 Czech Republic) and Roman PANTŮČEK (203 Czech Republic, belonging to the institution)
Edition
Phages 2011 - Bacteriophage Applications, 19-21 September 2011, St Hilda's College, Oxford, UK, 2011
Other information
Language
English
Type of outcome
Konferenční abstrakt
Field of Study
Genetics and molecular biology
Country of publisher
United Kingdom of Great Britain and Northern Ireland
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
RIV identification code
RIV/00216224:14310/11:00050057
Organization unit
Faculty of Science
Keywords in English
bacteriophage therapy; Staphylococcus aureus; bacteriophage endolysin; SH3b domain
Tags
International impact, Reviewed
Změněno: 14/10/2011 12:57, prof. RNDr. Roman Pantůček, Ph.D.
Abstract
V originále
In this study we focused on endolysin of staphylococcal bacteriophage 812, a member of unclassified SPO1-like viruses from family Myoviridae. The endolysin of phage 812 includes three domains: two catalytic domains and the SH3b-domain that is probably a cell wall targeting domain, which could be responsible for binding of endolysin to peptidoglycan. The gene sequence for the C-terminal SH3b domain was cloned in pET28 vector and expressed as a soluble protein in E. coli BL21 (DE3) to determine the 3D structure of the protein. A native variant of the SH3b protein was purified without the his-tag. Subsequently, three variants of this protein have been prepared: (i) non-labeled, (ii) single-labeled (15N) and (iii) double-labeled (13C, 15N) domain. The protein was purified to homogeneity using ammonium sulphate precipitation, anion exchange chromatography, cation exchange chromatography, and gel filtration. Functionality of the cell wall binding of the purified protein has been verified by a co-sedimentation test in using S. aureus peptidoglycan.
Links
GAP206/11/0758, research and development project |
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GA310/09/0459, research and development project |
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GP204/02/D099, research and development project |
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MSM0021622415, plan (intention) |
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TA01010405, research and development project |
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