Publications
131 results found
FAIRWEATHER N, BELL C, COCHRANE S, et al., 1994, MUTATIONS IN THE CONNEXIN-32 GENE IN X-LINKED DOMINANT CHARCOT-MARIE-TOOTH DISEASE (CMTX1), HUMAN MOLECULAR GENETICS, Vol: 3, Pages: 29-34, ISSN: 0964-6906
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- Citations: 182
DOUCE G, CROPLEY I, ROBERTS M, et al., 1994, A ROLE FOR TOXINS IN THE DEVELOPMENT OF ORAL VACCINES, 6th European Workshop on Bacterial Protein Toxins, Publisher: GUSTAV FISCHER VERLAG, Pages: 474-483, ISSN: 0172-5629
Makoff AJ, Charles IG, Fairweather NF, 1994, Recombinant antigens as components of a diphtheria-tetanus-pertussis vaccine., Bioprocess Technol, Vol: 19, Pages: 205-231, ISSN: 0888-7470
CHARLES I, RODGERS B, MUSGRAVE S, et al., 1993, EXPRESSION OF P.69/PERTACTIN FROM BORDETELLA-PERTUSSIS IN A BACULOVIRUS/INSECT CELL EXPRESSION SYSTEM - PROTECTIVE PROPERTIES OF THE RECOMNBINANT PROTEIN, RESEARCH IN MICROBIOLOGY, Vol: 144, Pages: 681-690, ISSN: 0923-2508
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- Citations: 6
FAIRWEATHER NF, SANDERS D, SLATER D, et 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
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- Citations: 15
ANDERSON MD, FAIRWEATHER N, CHARLES IG, et al., 1993, CRYSTALLOGRAPHIC CHARACTERIZATION OF TETANUS TOXIN FRAGMENT-C, JOURNAL OF MOLECULAR BIOLOGY, Vol: 230, Pages: 673-674, ISSN: 0022-2836
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- Citations: 9
HOGY B, FAIRWEATHER N, LINDER D, et 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
WELLER U, FAIRWEATHER N, SANDERS D, et 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
STRUGNELL R, DOUGAN G, CHATFIELD S, et 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
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- Citations: 91
CHATFIELD SN, CHARLES IG, MAKOFF AJ, et 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
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- Citations: 191
Li J, Fairweather NF, Novotny P, et 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.
LI JL, FAIRWEATHER NF, NOVOTNY P, et 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
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- Citations: 38
CHATFIELD SN, FAIRWEATHER N, CHARLES I, et 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
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- Citations: 65
ROBERTS M, TITE JP, FAIRWEATHER NF, et al., 1992, RECOMBINANT P69/PERTACTIN - IMMUNOGENICITY AND PROTECTION OF MICE AGAINST BORDETELLA-PERTUSSIS INFECTION, VACCINE, Vol: 10, Pages: 43-52, ISSN: 0264-410X
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- Citations: 30
Roberts M, Tite JP, Fairweather NF, et 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.
ROMANOS MA, CLARE JJ, BEESLEY KM, et 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
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- Citations: 81
LIPSCOMBE M, CHARLES IG, ROBERTS M, et 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
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- Citations: 56
ROBERTS M, FAIRWEATHER NF, LEININGER E, et 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
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- Citations: 80
CHARLES IG, LI JL, ROBERTS M, et 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
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- Citations: 52
CHARLES IG, RODGERS BC, MAKOFF AJ, et 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
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- Citations: 26
ROMANOS MA, MAKOFF AJ, FAIRWEATHER NF, et 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
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- Citations: 77
LEININGER E, ROBERTS M, KENIMER JG, et 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
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- Citations: 224
SERVOS S, CHATFIELD S, HONE D, et 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
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- Citations: 23
Makoff AJ, Oxer MD, Ballantine SP, et 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.
YANG DM, FAIRWEATHER N, BUTTON LL, et 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
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- Citations: 174
FAIRWEATHER NF, CHATFIELD SN, CHARLES IG, et al., 1990, USE OF LIVE ATTENUATED BACTERIA TO STIMULATE IMMUNITY, RESEARCH IN MICROBIOLOGY, Vol: 141, Pages: 769-773, ISSN: 0923-2508
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- Citations: 15
FAIRWEATHER NF, CHATFIELD SN, MAKOFF AJ, et 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
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- Citations: 94
STRUGNELL RA, MASKELL D, FAIRWEATHER N, et al., 1990, STABLE EXPRESSION OF FOREIGN ANTIGENS FROM THE CHROMOSOME OF SALMONELLA-TYPHIMURIUM VACCINE STRAINS, GENE, Vol: 88, Pages: 57-63, ISSN: 0378-1119
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- Citations: 63
MAKOFF AJ, OXER MD, ROMANOS MA, et 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
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- Citations: 135
Makoff AJ, Oxer MD, Romanos MA, et 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.
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