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Cloning, expression, crystallization and preliminary X‐ray diffraction studies of staphylococcal superantigen‐like protein 1 (SSL1)
Author(s) -
Dutta Debabrata,
Dutta Anirudha,
Bhattacharjee Atanu,
Basak Amit,
Das Amit Kumar
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14006967
Subject(s) - superantigen , polyethylene glycol , crystallization , staphylococcus aureus , microbiology and biotechnology , escherichia coli , cloning (programming) , enterotoxin , chemistry , recombinant dna , expression vector , crystallography , biology , gene , bacteria , biochemistry , genetics , organic chemistry , computer science , programming language
Staphylococcus aureus produces a family of exotoxins which are structural homologues of superantigens and thus are called staphylococcal superantigen‐like proteins (SSLs). Amongst the 14 SSL genes, ssl1 (SAOUHSC_00383) has been cloned in the pQE30 expression vector, overexpressed in Escherichia coli M15 (pREP4) cells and the protein purified to homogeneity. The protein was crystallized using 6% Tacsimate pH 6.0, 0.1 M MES pH 6.0, 25%( w / v ) polyethylene glycol 3350, 100 m M NDSB 256 at 298 K by the sitting‐drop vapour‐diffusion method. The crystals belonged to space group P 2 1 , with unit‐cell parameters a = 77.9, b = 70.5, c = 126.5 Å, β = 106.2°. X‐ray diffraction data were collected and processed to a maximum resolution of 2.5 Å. The crystal contains six molecules in the asymmetric unit.