Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F11%3A00050057" target="_blank" >RIV/00216224:14310/11:00050057 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin

Popis výsledku v původním jazyce

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 acell 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.

Název v anglickém jazyce

Cloning, expression and structure determination of SH3b cell wall binding domain of bacteriophage 812 endolysin

Popis výsledku anglicky

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 acell 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.

Druh

O - Ostatní výsledky

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2011

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů