title: Cloning, sequencing and expression of porcine CD40 ligand in Escherichia coli and human and porcine cells.
abstract: The CD40L ligand (CD40L) plays an important role in the interaction between antigen-specific T lymphocytes and antigen-presenting cells. The porcine CD40L encoding gene was isolated from porcine peripheral blood mononuclear cells (PBMC) using RT-PCR. Sequence analysis of the cloned CD40L gene showed an open reading frame of 786 base pairs encoding a 262 amino acid protein with a predicted molecular mass of 29 kD. The deduced amino acid sequence of the porcine CD40L shared 82%, 88% and 93% similarity with the CD40L protein of mouse, human and cattle. The isolated CD40L sequence was expressed as a hexahistidine fusion protein in Escherichia coli and purified by affinity chromatography. The analysis of the CD40L-expression in human 293 and porcine MAX cells by immunofluorescence showed its location on the cell surface.
authors.affiliation: Federal Research Center for Virus Diseases of Animals, Institute for Immunology, Tübingen, Germany. daniel.wienhold@tue.bfav.de
authors.forename: Daniel
authors.initials: D
authors.lastname: Wienhold
authors.forename: Nicola
authors.initials: N
authors.lastname: Berger
authors.forename: Elisenda
authors.initials: E
authors.lastname: Armengol
authors.forename: Mathias
authors.initials: M
authors.lastname: Büttner
authors.forename: Armin
authors.initials: A
authors.lastname: Saalmüller
authors.forename: Eberhard
authors.initials: E
authors.lastname: Pfaff
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D023201:CD40 Ligand / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008969:Molecular Sequence Data; D013552:Swine / Q000378:metabolism*
chemical_list: D023201:CD40 Ligand