title: Vectors and P64k gene targeting for tandem affinity purification in Neisseria meningitidis.
abstract: We present and describe the construction of tagging cassettes and plasmids for tandem affinity purification (TAP) of proteins in Neisseria meningitidis. The tagging cassette is designed for carboxyl-terminal tagging of proteins and it contains only two repeats of IgG-binding units. P64k protein from N. meningitidis was chosen to fuse at these new affinity tags. This protein is well recognized in immunoassays by serum from human convalescent meningococcal disease and it is highly immunogenic in animals. To continue the characterization of this meningococcal antigen, we designed and constructed two vectors for use in TAP purification method. We also carried-out preliminary test to check the correct expression of the protein fused in these vectors.
authors.affiliation: Universidad Politecnica de Catalunya, Ingenieria Quimica, Colom 1. EUETIT. TR1, 08222 Terrassa, Spain. dasiel.borroto@upc.es
authors.forename: Dasiel Oscar
authors.initials: DO
authors.lastname: Borroto Escuela
authors.forename: Mileydis
authors.initials: M
authors.lastname: Perez Alea
authors.forename: Wilber Romero
authors.initials: WR
authors.lastname: Fernandez
authors.forename: Daniel Bello
authors.initials: DB
authors.lastname: Gil
mesh_terms: D000345:Affinity Labels / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016254:Mutagenesis, Insertional / Q000379:methods; D009345:Neisseria meningitidis / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D000345:Affinity Labels; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011994:Recombinant Proteins; C112623:lpdA protein, Neisseria meningitidis