title: Purification of recombinantly expressed human cluster determinant 4 cytoplasmic domain.
abstract: A DNA fragment coding for the human CD4 cytoplasmic domain (residues 394-433) was cloned into the pET15b expression vector. The resulting plasmid was used for synthesis of the polyhistidine-tagged 5.10(3) M(r) CD4 peptide in Escherichia coli BL21(DE3)Star. The CD4 cytoplasmic domain was purified under denaturing and reducing conditions by a two-step procedure using immobilized metal affinity chromatography and gel permeation chromatography. The purified CD4 cytoplasmic domain is soluble and functional without any specific refolding steps. The yield of the described purification procedure was approximately 5 mg peptide per liter culture volume.
authors.affiliation: Institut für Molekulare Biotechnologie, Beutenbergstr. 11, 07745, Jena, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Preusser
authors.forename: Gesa
authors.initials: G
authors.lastname: Jonas
authors.forename: Dieter
authors.initials: D
authors.lastname: Willbold
mesh_terms: D001483:Base Sequence; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008562:Membrane Glycoproteins / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins