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  • 1
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 15 (1995), S. 97-106 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The 5C outer membrane protein, one of the N. meningitidis class 5 proteins, was preferably expressed in bacteria isolated from the nasopharynx and its role in adhering to the mucosal cells and invading them as well as the development of anti-5C antibodies in healthy carriers was demonstrated. Anti-5C monoclonal antibodies are bactericidal in the presence of the human complement. The immunodominant region of the 5C protein is highly conserved among the different strains of N. meningitidis, and the opc gene, which encodes the protein, does not seem to show antigenic variations.Here the isolation of the opc gene from the Cuban strain B:4:P1.15 by PCR (Polymerase Chain Reaction) is presented. Under the regulation of the tryptophan promoter, the gene was cloned and sequenced in E. coli with a high level of expression and fused to the amino-terminal end of the interleukin-2 gene. In the dot-blot experiments, the presence of the gene in those strains which did not express the protein in the whole cell ELISA was also detectable.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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  • 2
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The gene coding for the class 1 outer membrane protein from the Neisseria meningitidis strain B385 (B : 4 : P1.15) was isolated by the Polymerase Chain Reaction (PCR) and cloned into the Sma I cut M13mp 18 vector. Then, a Xba I restriction site was created by using an oligonucleotide-directed in vitro mutagenesis system at the start of the coding fragment. In order to express the protein, this fragment was fused to 180 base pairs corresponding to 60 amino acids from the N terminus of interleukin-2 under the control of the tryptophan promoter (Ptrp). The expression was confirmed by Western-blotting where the recombinant protein (PILM28) was detected by bactericidal monoclonal antibodies (MABs). The recombinant polypeptide was partially purified and used to elicit murine antibodies, able to recognize the meningococcal class 1 subtype 15.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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