Expression in Escherichia coli of the lpdA gene, protein sequence analysis and immunological characterization of the P64k protein from Neisseria meningitidis.
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.
Division of Vaccines, Centro de Ingeniería Genética y Biotecnología, La Habana, Cuba.
G
G
Guillén
A
A
Alvarez
R
R
Silva
V
V
Morera
S
S
González
A
A
Musacchio
V
V
Besada
E
E
Coizeau
E
E
Caballero
C
C
Nazabal
T
T
Carmenate
L J
LJ
González
R
R
Estrada
Y
Y
Támbara
G
G
Padrón
L
L
Herrera
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli; D015870:Gene Expression; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis; D011994:Recombinant Proteins / Q000096:biosynthesis; D017422:Sequence Analysis, DNA
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D011994:Recombinant Proteins; C112623:lpdA protein, Neisseria meningitidis