About: Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin     Goto   Sponge   NotDistinct   Permalink

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  • 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.
  • 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. (en)
Title
  • Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin
  • Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin (en)
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  • Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin
  • Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin (en)
skos:notation
  • RIV/00216224:14310/11:00050057!RIV12-TA0-14310___
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  • P(GA310/09/0459), P(GAP206/11/0758), P(GP204/02/D099), P(TA01010405), Z(MSM0021622415)
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  • 190638
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  • RIV/00216224:14310/11:00050057
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  • bacteriophage therapy; Staphylococcus aureus; bacteriophage endolysin; SH3b domain (en)
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  • [AA634D126475]
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  • Benešík, Martin
  • Dopitová, Radka
  • Doškař, Jiří
  • Janda, Lubomír
  • Moša, Marek
  • Nováček, Jiří
  • Pantůček, Roman
  • Růžičková, Vladislava
  • Žídek, Lukáš
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  • 14310
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