Vectors and P64k gene targeting for tandem affinity purification in Neisseria meningitidis.
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.
Borroto Escuela
Dasiel Oscar
DO
Universidad Politecnica de Catalunya, Ingenieria Quimica, Colom 1. EUETIT. TR1, 08222 Terrassa, Spain. dasiel.borroto@upc.es
Perez Alea
Mileydis
M
Fernandez
Wilber Romero
WR
Gil
Daniel Bello
DB
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
D000345:Affinity Labels; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011994:Recombinant Proteins; C112623:lpdA protein, Neisseria meningitidis