Imperial College London

professor neil fairweather

Faculty of Natural SciencesDepartment of Life Sciences

Professor of Microbiology
 
 
 
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Contact

 

+44 (0)20 7594 5247n.fairweather

 
 
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Location

 

1.44Flowers buildingSouth Kensington Campus

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Summary

 

Publications

Publication Type
Year
to

125 results found

FAIRWEATHER NF, SANDERS D, SLATER D, HUDEL M, HABERMANN E, WELLER Uet al., 1993, PRODUCTION OF BIOLOGICALLY-ACTIVE LIGHT-CHAIN OF TETANUS TOXIN IN ESCHERICHIA-COLI - EVIDENCE FOR THE IMPORTANCE OF THE C-TERMINAL 16 AMINO-ACIDS FOR FULL BIOLOGICAL-ACTIVITY, FEBS LETTERS, Vol: 323, Pages: 218-222, ISSN: 0014-5793

JOURNAL ARTICLE

HOGY B, FAIRWEATHER N, LINDER D, WELLER U, HABERMANN Eet al., 1993, THE GANGLIOSIDE BINDING MOIETY OF TETANUS TOXIN - EXPRESSION, FRAGMENTATION AND RETROGRADE AXONAL-TRANSPORT, 5TH EUROPEAN WORKSHOP ON BACTERIAL PROTEIN TOXINS, Publisher: GUSTAV FISCHER VERLAG, Pages: 156-157, ISSN: 0172-5629

CONFERENCE PAPER

WELLER U, FAIRWEATHER N, SANDERS D, AHNERTHILGER G, PAYNE M, HABERMANN Eet al., 1993, EXPRESSION OF TETANUS TOXIN LIGHT CHAIN AND MUTANTS OF CYS-438 IN ESCHERICHIA-COLI - PURIFICATION AND BIOLOGICAL-ACTIVITIES OF THE RECOMBINANT PROTEINS, 5TH EUROPEAN WORKSHOP ON BACTERIAL PROTEIN TOXINS, Publisher: GUSTAV FISCHER VERLAG, Pages: 99-100, ISSN: 0172-5629

CONFERENCE PAPER

CHATFIELD SN, CHARLES IG, MAKOFF AJ, OXER MD, DOUGAN G, PICKARD D, SLATER D, FAIRWEATHER NFet al., 1992, USE OF THE NIRB PROMOTER TO DIRECT THE STABLE EXPRESSION OF HETEROLOGOUS ANTIGENS IN SALMONELLA ORAL VACCINE STRAINS - DEVELOPMENT OF A SINGLE-DOSE ORAL TETANUS VACCINE, BIO-TECHNOLOGY, Vol: 10, Pages: 888-892, ISSN: 0733-222X

JOURNAL ARTICLE

CHATFIELD SN, FAIRWEATHER N, CHARLES I, PICKARD D, LEVINE M, HONE D, POSADA M, STRUGNELL RA, DOUGAN Get al., 1992, CONSTRUCTION OF A GENETICALLY DEFINED SALMONELLA-TYPHI TY2 AROA, AROC MUTANT FOR THE ENGINEERING OF A CANDIDATE ORAL TYPHOID TETANUS VACCINE, VACCINE, Vol: 10, Pages: 53-60, ISSN: 0264-410X

JOURNAL ARTICLE

LI JL, FAIRWEATHER NF, NOVOTNY P, DOUGAN G, CHARLES IGet al., 1992, CLONING, NUCLEOTIDE-SEQUENCE AND HETEROLOGOUS EXPRESSION OF THE PROTECTIVE OUTER-MEMBRANE PROTEIN P.68 PERTACTIN FROM BORDETELLA-BRONCHISEPTICA, JOURNAL OF GENERAL MICROBIOLOGY, Vol: 138, Pages: 1697-1705, ISSN: 0022-1287

JOURNAL ARTICLE

Li J, Fairweather NF, Novotny P, Dougan G, Charles IGet al., 1992, Cloning, nucleotide sequence and heterologous expression of the protective outer-membrane protein P.68 pertactin from Bordetella bronchiseptica., J Gen Microbiol, Vol: 138 Pt 8, Pages: 1697-1705, ISSN: 0022-1287

The prn gene encoding the 68 kDa protective outer-membrane protein of Bordetella bronchiseptica (P.68 pertactin) was cloned, sequenced and expressed in Escherichia coli. The gene was isolated by DNA:DNA hybridization experiments using a radioactively-labelled fragment of the homologous prn gene from Bordetella parapertussis. DNA sequence analysis reveals that the gene is capable of encoding a protein with a molecular mass of 93996 Da (P.94); this precursor molecule is processed to form the P.68 antigen on the surface of B. bronchiseptica. Heterologous expression of the full-length gene encoding P.94 in Escherichia coli results in similar processing, with the P.68 antigen targeted to the bacterial outer membrane. Comparison of P.94 with the P.93 and P.95 precursors, encoding homologous proteins from Bordetella pertussis and B. parapertussis, shows a high degree (greater than 90%) of homology. The major differences between all three proteins occur in the number of repeats of the two families (Gly-Gly-Xaa-Xaa-Pro)n and (Pro-Gln-Pro)n of reiterated sequence motifs.

JOURNAL ARTICLE

ROBERTS M, TITE JP, FAIRWEATHER NF, DOUGAN G, CHARLES IGet al., 1992, RECOMBINANT P69/PERTACTIN - IMMUNOGENICITY AND PROTECTION OF MICE AGAINST BORDETELLA-PERTUSSIS INFECTION, VACCINE, Vol: 10, Pages: 43-52, ISSN: 0264-410X

JOURNAL ARTICLE

Roberts M, Tite JP, Fairweather NF, Dougan G, Charles IGet al., 1992, Recombinant P.69/pertactin: immunogenicity and protection of mice against Bordetella pertussis infection., Vaccine, Vol: 10, Pages: 43-48, ISSN: 0264-410X

The immunogenicity of recombinant (r-) pertactin was examined. Parenteral immunization of mice with natural or r-pertactin produced a similar increase in serum anti-pertactin antibodies and a decrease in Bordetella pertussis lung counts following aerosol challenge. Study of the kinetics of B. pertussis growth in the respiratory tract of immunized and control mice revealed that immunization with r-pertactin halted the multiplication of B. pertussis in the lungs and facilitated the early onset of bacterial clearance. In the trachea, bacterial numbers declined sharply in immunized animals during the first 3 days after challenge but thereafter B. pertussis numbers remained fairly constant throughout the rest of the experiment. Very low doses (0.1 micrograms) of r-pertactin were immunogenic and protective but only if the antigen was absorbed to alhydrogel. In vitro proliferation assays with lymphocytes from mice primed with either natural or r-pertactin indicated that the major T-cell epitopes of pertactin are conserved in the recombinant protein.

JOURNAL ARTICLE

STRUGNELL R, DOUGAN G, CHATFIELD S, CHARLES I, FAIRWEATHER N, TITE J, LI JL, BEESLEY J, ROBERTS Met al., 1992, CHARACTERIZATION OF A SALMONELLA-TYPHIMURIUM-ARO VACCINE STRAIN EXPRESSING THE P.69 ANTIGEN OF BORDETELLA-PERTUSSIS, INFECTION AND IMMUNITY, Vol: 60, Pages: 3994-4002, ISSN: 0019-9567

JOURNAL ARTICLE

CHARLES IG, LI JL, ROBERTS M, BEESLEY K, ROMANOS M, PICKARD DJ, FRANCIS M, CAMPBELL D, DOUGAN G, BRENNAN MJ, MANCLARK CR, JENSEN MA, HERON I, CHUBB A, NOVOTNY P, FAIRWEATHER NFet al., 1991, IDENTIFICATION AND CHARACTERIZATION OF A PROTECTIVE IMMUNODOMINANT B-CELL EPITOPE OF PERTACTIN (P.69) FROM BORDETELLA-PERTUSSIS, EUROPEAN JOURNAL OF IMMUNOLOGY, Vol: 21, Pages: 1147-1153, ISSN: 0014-2980

JOURNAL ARTICLE

CHARLES IG, RODGERS BC, MAKOFF AJ, CHATFIELD SN, SLATER DE, FAIRWEATHER NFet al., 1991, SYNTHESIS OF TETANUS TOXIN FRAGMENT-C IN INSECT CELLS BY USE OF A BACULOVIRUS EXPRESSION SYSTEM, INFECTION AND IMMUNITY, Vol: 59, Pages: 1627-1632, ISSN: 0019-9567

JOURNAL ARTICLE

LEININGER E, ROBERTS M, KENIMER JG, CHARLES IG, FAIRWEATHER N, NOVOTNY P, BRENNAN MJet al., 1991, PERTACTIN, AN ARG-GLY-ASP-CONTAINING BORDETELLA-PERTUSSIS SURFACE PROTEIN THAT PROMOTES ADHERENCE OF MAMMALIAN-CELLS, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol: 88, Pages: 345-349, ISSN: 0027-8424

JOURNAL ARTICLE

LIPSCOMBE M, CHARLES IG, ROBERTS M, DOUGAN G, TITE J, FAIRWEATHER NFet al., 1991, INTRANASAL IMMUNIZATION USING THE B-SUBUNIT OF THE ESCHERICHIA-COLI HEAT-LABILE TOXIN FUSED TO AN EPITOPE OF THE BORDETELLA-PERTUSSIS P69 ANTIGEN, MOLECULAR MICROBIOLOGY, Vol: 5, Pages: 1385-1392, ISSN: 0950-382X

JOURNAL ARTICLE

ROBERTS M, FAIRWEATHER NF, LEININGER E, PICKARD D, HEWLETT EL, ROBINSON A, HAYWARD C, DOUGAN G, CHARLES IGet al., 1991, CONSTRUCTION AND CHARACTERIZATION OF BORDETELLA-PERTUSSIS MUTANTS LACKING THE VIR-REGULATED P69 OUTER-MEMBRANE PROTEIN, MOLECULAR MICROBIOLOGY, Vol: 5, Pages: 1393-1404, ISSN: 0950-382X

JOURNAL ARTICLE

ROMANOS MA, CLARE JJ, BEESLEY KM, RAYMENT FB, BALLANTINE SP, MAKOFF AJ, DOUGAN G, FAIRWEATHER NF, CHARLES IGet al., 1991, RECOMBINANT BORDETELLA-PERTUSSIS PERTACTIN (P69) FROM THE YEAST PICHIA-PASTORIS - HIGH-LEVEL PRODUCTION AND IMMUNOLOGICAL PROPERTIES, VACCINE, Vol: 9, Pages: 901-906, ISSN: 0264-410X

JOURNAL ARTICLE

ROMANOS MA, MAKOFF AJ, FAIRWEATHER NF, BEESLEY KM, SLATER DE, RAYMENT FB, PAYNE MM, CLARE JJet al., 1991, EXPRESSION OF TETANUS TOXIN FRAGMENT-C IN YEAST - GENE SYNTHESIS IS REQUIRED TO ELIMINATE FORTUITOUS POLYADENYLATION SITES IN AT-RICH DNA, NUCLEIC ACIDS RESEARCH, Vol: 19, Pages: 1461-1467, ISSN: 0305-1048

JOURNAL ARTICLE

SERVOS S, CHATFIELD S, HONE D, LEVINE M, DIMITRIADIS G, PICKARD D, DOUGAN G, FAIRWEATHER N, CHARLES Iet al., 1991, MOLECULAR-CLONING AND CHARACTERIZATION OF THE AROD GENE ENCODING 3-DEHYDROQUINASE FROM SALMONELLA-TYPHI, JOURNAL OF GENERAL MICROBIOLOGY, Vol: 137, Pages: 147-152, ISSN: 0022-1287

JOURNAL ARTICLE

FAIRWEATHER NF, CHATFIELD SN, CHARLES IG, ROBERTS M, LIPSCOMBE M, LI LJ, STRUGNELL D, COMERFORD S, TITE J, DOUGAN Get al., 1990, USE OF LIVE ATTENUATED BACTERIA TO STIMULATE IMMUNITY, RESEARCH IN MICROBIOLOGY, Vol: 141, Pages: 769-773, ISSN: 0923-2508

JOURNAL ARTICLE

FAIRWEATHER NF, CHATFIELD SN, MAKOFF AJ, STRUGNELL RA, BESTER J, MASKELL DJ, DOUGAN Get al., 1990, ORAL VACCINATION OF MICE AGAINST TETANUS BY USE OF A LIVE ATTENUATED SALMONELLA CARRIER, INFECTION AND IMMUNITY, Vol: 58, Pages: 1323-1326, ISSN: 0019-9567

JOURNAL ARTICLE

Makoff AJ, Oxer MD, Ballantine SP, Fairweather NF, Charles IGet al., 1990, Protective surface antigen P69 of Bordetella pertussis: its characterization and very high level expression in Escherichia coli., Biotechnology (N Y), Vol: 8, Pages: 1030-1033, ISSN: 0733-222X

The surface antigen, P69 of Bordetella pertussis, an N-terminal fragment of the precursor protein, P93, is likely to be an important component of future subunit vaccines against whooping cough. We have expressed several defined N-terminal fragments of P93 in E. coli and compared their electrophoretic mobilities with that of purified P69 from B. pertussis. These experiments show that P69 is considerably smaller than the 69 kD originally estimated from its gel mobility and is probably 60.4 kD in size. Our initial plasmids expressed only very low levels of this antigen. We diagnosed the limiting factor to be a poor ribosome binding site (RBS) by demonstrating a large stimulation of expression on a two-cistron plasmid. The limitation of expression could be completely overcome by only two base changes close to the initiation codon, resulting in a further increase in expression of P69 at levels to 30-40% total cell protein. Although the protein accumulated as insoluble inclusion bodies, it could be solubilized by guanidinium chloride.

JOURNAL ARTICLE

STRUGNELL RA, MASKELL D, FAIRWEATHER N, PICKARD D, COCKAYNE A, PENN C, DOUGAN Get al., 1990, STABLE EXPRESSION OF FOREIGN ANTIGENS FROM THE CHROMOSOME OF SALMONELLA-TYPHIMURIUM VACCINE STRAINS, GENE, Vol: 88, Pages: 57-63, ISSN: 0378-1119

JOURNAL ARTICLE

YANG DM, FAIRWEATHER N, BUTTON LL, MCMASTER WR, KAHL LP, LIEW FYet al., 1990, ORAL SALMONELLA-TYPHIMURIUM (AROA-) VACCINE EXPRESSING A MAJOR LEISHMANIAL SURFACE PROTEIN (GP63) PREFERENTIALLY INDUCES T-HELPER 1-CELLS AND PROTECTIVE IMMUNITY AGAINST LEISHMANIASIS, JOURNAL OF IMMUNOLOGY, Vol: 145, Pages: 2281-2285, ISSN: 0022-1767

JOURNAL ARTICLE

CHARLES IG, DOUGAN G, PICKARD D, CHATFIELD S, SMITH M, NOVOTNY P, MORRISSEY P, FAIRWEATHER NFet al., 1989, MOLECULAR-CLONING AND CHARACTERIZATION OF PROTECTIVE OUTER-MEMBRANE PROTEIN-P.69 FROM BORDETELLA-PERTUSSIS, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol: 86, Pages: 3554-3558, ISSN: 0027-8424

JOURNAL ARTICLE

LESLIE D, FAIRWEATHER N, PICKARD D, DOUGAN G, KEHOE Met al., 1989, PHOSPHOLIPASE-C AND HEMOLYTIC ACTIVITIES OF CLOSTRIDIUM-PERFRINGENS ALPHA-TOXIN CLONED IN ESCHERICHIA-COLI - SEQUENCE AND HOMOLOGY WITH A BACILLUS-CEREUS PHOSPHOLIPASE-C, MOLECULAR MICROBIOLOGY, Vol: 3, Pages: 383-392, ISSN: 0950-382X

JOURNAL ARTICLE

MAKOFF AJ, BALLANTINE SP, SMALLWOOD AE, FAIRWEATHER NFet al., 1989, EXPRESSION OF TETANUS TOXIN FRAGMENT-C IN ESCHERICHIA-COLI - ITS PURIFICATION AND POTENTIAL USE AS A VACCINE, BIO-TECHNOLOGY, Vol: 7, Pages: 1043-1046, ISSN: 0733-222X

JOURNAL ARTICLE

MAKOFF AJ, OXER MD, ROMANOS MA, FAIRWEATHER NF, BALLANTINE Set al., 1989, EXPRESSION OF TETANUS TOXIN FRAGMENT-C IN ESCHERICHIA-COLI - HIGH-LEVEL EXPRESSION BY REMOVING RARE CODONS, NUCLEIC ACIDS RESEARCH, Vol: 17, Pages: 10191-10202, ISSN: 0305-1048

JOURNAL ARTICLE

Makoff AJ, Oxer MD, Romanos MA, Fairweather NF, Ballantine Set al., 1989, Expression of tetanus toxin fragment C in E. coli: high level expression by removing rare codons., Nucleic Acids Res, Vol: 17, Pages: 10191-10202, ISSN: 0305-1048

Tetanus toxin fragment C had been previously expressed in Escherichia coli at 3-4% cell protein. The codon bias for tetanus toxin in Clostridium tetani is very different from that of highly expressed homologous genes in E. coli, resulting in the presence of many rare E. coli codons in the sequence encoding fragment C. We have replaced the coding sequence by sequence optimized for codon usage in E. coli, and show that the expression of fragment C is increased. Although the level of mRNA also increased this appeared to be a secondary consequence of more efficient translation. Complete sequence replacement increased expression to approximately 11-14% cell protein but only after the promoter strength had been improved.

JOURNAL ARTICLE

Charles IG, Dougan G, Pickard D, Charfield S, Smith M, Novotny P, Fairweather Net al., 1988, Molecular cloning and analysis of P. 69, a vir-controlled protein from Bordetella pertussis., Tokai J Exp Clin Med, Vol: 13 Suppl, Pages: 227-234, ISSN: 0385-0005

The cloning sequencing and analysis of an important antigenic component of Bordetella pertussis is described. The gene for P.69, in common with a variety of other so called "virulence" genes, (e.g., adenylate cyclase (AC), pertussis toxin (PT) and filamentous haemagglutinin (FHA)), is under control of the vir locus. The protein P.69 is externally localised on cells and protein preparations are protective as judged by the mouse intra-cerebral challenge test. The gene encoding the P.69 antigen was isolated by hybridization of mixed oligonucleotide probes against B. pertussis genomic DNA. These oligonucleotides were designed from the protein sequence data obtained from a cyanogen bromide digest of the P.69 protein. DNA sequence analysis reveals a G:C rich gene capable of encoding a protein of 910 amino acids and Mr of 93478, whose likely promoter and ribosome binding sites show little homology to their E.coli counterpart. In common with some of the genes in the PT operon the sequence 5'-CCTGG-3' was found 5' to the ATG initiation codon. At the 3', end 29 bases after the TAA stop codon, the sequence 5'GTTTTTCCT-3' was found in an equivalent position to the same sequence in the PT operon. Examination of the protein sequence reveals two regions with directly repeated elements (GGAVP)3(GGFGP)2 and (PQP)5.

JOURNAL ARTICLE

FAIRWEATHER NF, LYNESS VA, MASKELL DJ, 1987, IMMUNIZATION OF MICE AGAINST TETANUS WITH FRAGMENTS OF TETANUS TOXIN SYNTHESIZED IN ESCHERICHIA-COLI, INFECTION AND IMMUNITY, Vol: 55, Pages: 2541-2545, ISSN: 0019-9567

JOURNAL ARTICLE

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