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Cloning, expression, purification, crystallization and preliminary X-ray diffraction studies of a major group 7 allergen, Der f 7, from the dust mite Dermatophagoides farinae.
- Source :
-
Acta crystallographica. Section F, Structural biology and crystallization communications [Acta Crystallogr Sect F Struct Biol Cryst Commun] 2011 Dec 01; Vol. 67 (Pt 12), pp. 1612-5. Date of Electronic Publication: 2011 Nov 26. - Publication Year :
- 2011
-
Abstract
- Der f 7 is a major group 7 allergen from the dust mite Dermatophagoides farinae that shows 86% sequence identity to the homologous allergen Der p 7 from D. pteronyssinus. Der f 7 was successfully overexpressed in an Escherichia coli expression system and purified to homogeneity using Ni-NTA affinity and size-exclusion column chromatography. SeMet-labelled Der f 7 was crystallized by the hanging-drop vapour-diffusion method using a reservoir solution consisting of 0.1 M bis-tris pH 7.4 and 28% polyethylene glycol monomethyl ether 2000 at 293 K. X-ray diffraction data were collected to 2.24 Å resolution using synchrotron radiation. The crystals belonged to the orthorhombic system, space group P2(1)2(1)2(1), with unit-cell parameters a = 50.19, b = 58.67, c = 123.81 Å. Based on the estimated Matthews coefficient (2.16 Å(3) Da(-1)), two molecules of Der f 7 could be present in the asymmetric unit of the crystal lattice.
- Subjects :
- Animals
Antigens, Dermatophagoides genetics
Antigens, Dermatophagoides isolation & purification
Arthropod Proteins genetics
Arthropod Proteins isolation & purification
Cloning, Molecular
Crystallization
Crystallography, X-Ray
Gene Expression
Antigens, Dermatophagoides chemistry
Arthropod Proteins chemistry
Dermatophagoides farinae chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1744-3091
- Volume :
- 67
- Issue :
- Pt 12
- Database :
- MEDLINE
- Journal :
- Acta crystallographica. Section F, Structural biology and crystallization communications
- Publication Type :
- Academic Journal
- Accession number :
- 22139179
- Full Text :
- https://doi.org/10.1107/S174430911103836X