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Biotechnology and Applied Biochemistry (1998) 27, (189–196) (Printed in Great Britain)

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Expression in Escherichia coli of the lpdA gene, protein sequence analysis and immunological characterization of the P64k protein from Neisseria meningitidis

Gerardo Guillén*, Anabel Alvarez*, Ricardo Silva*, Vivian Morera†, Sonia González*, Alexis Musacchio*, Vladimir Besada†, Edelgis Coizeau*, Evelin Caballero*, Consuelo Nazabal*, Tania Carmenate*, Luis J. González†, Regla Estrada†, Yanet Támbara†, Gabriel Padrón† and Luis Herrera*

*Division of Vaccines, Centro de Ingeniería Genética y Biotecnología, Apartado 6162, La Habana, Cuba

Abbreviations: LDH, dihydrolipoamide dehydrogenase; mAbs monoclonal antibodies; OMP, outer membrane protein; SCM, S-carboxymethyl.

Correspondence: Gerardo Guillén.

To whom correspondence should be addressed.

By making use of recombinant DNA technology it is possible to characterize meningococcal outer membrane proteins (OMPs) capable of stimulating a host immune response. The lpdA gene, which codes for an OMP (P64k) from Neisseria meningitidis, was cloned in Escherichia coli. The recombinant protein was recognized by sera from patients convalescing from meningococcal disease. The monoclonal antibodies obtained against the recombinant protein recognized the natural protein on a Western blot, and monoclonal antibody 114 was assayed in ELISA with a panel of 85 N. meningitidis strains. The protein was recognized in 81 strains (95.3%); the strains that were not recognized were neither epidemic nor isolated from systemic disease. The complete amino acid sequence of P64k was obtained by automatic sequencing and MS.

(Received 12 August 1997/30 October 1997; accepted 16 December 1997)

The Biochemical Society and the Medical Research Society © 1998