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Infection and Immunity logoLink to Infection and Immunity
. 1989 Apr;57(4):1347–1350. doi: 10.1128/iai.57.4.1347-1350.1989

Construction of a plasmid for expression of foreign epitopes as fusion proteins with subunit B of Escherichia coli heat-labile enterotoxin.

F Schödel 1, H Will 1
PMCID: PMC313276  PMID: 2647637

Abstract

A novel vector (pFS2.2) for high-level expression of fusion polypeptides with the nontoxic subunit B (LT-B) of Escherichia coli heat-labile enterotoxin in Escherichia coli and salmonellae is presented. It carries the complete coding sequence of LT-B under lac promoter control and a universal polylinker site for the in-frame insertion of foreign genes at the LT-B gene 3' end. By using this vector, fusion proteins comprising parts of the human or woodchuck hepatitis B virus surface and nucleocapsid antigens are expressed in E. coli and salmonella.

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Selected References

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  1. Beasley R. P., Hwang L. Y., Lin C. C., Chien C. S. Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22 707 men in Taiwan. Lancet. 1981 Nov 21;2(8256):1129–1133. doi: 10.1016/s0140-6736(81)90585-7. [DOI] [PubMed] [Google Scholar]
  2. Clements J. D., El-Morshidy S. Construction of a potential live oral bivalent vaccine for typhoid fever and cholera-Escherichia coli-related diarrheas. Infect Immun. 1984 Nov;46(2):564–569. doi: 10.1128/iai.46.2.564-569.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clements J. D., Flint D. C., Engert R. F., Klipstein F. A. Cloning and molecular characterization of the B subunit of Escherichia coli heat-labile enterotoxin. Infect Immun. 1983 May;40(2):653–658. doi: 10.1128/iai.40.2.653-658.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clements J. D., Lyon F. L., Lowe K. L., Farrand A. L., el-Morshidy S. Oral immunization of mice with attenuated Salmonella enteritidis containing a recombinant plasmid which codes for production of the B subunit of heat-labile Escherichia coli enterotoxin. Infect Immun. 1986 Sep;53(3):685–692. doi: 10.1128/iai.53.3.685-692.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dallas W. S., Falkow S. Amino acid sequence homology between cholera toxin and Escherichia coli heat-labile toxin. Nature. 1980 Dec 4;288(5790):499–501. doi: 10.1038/288499a0. [DOI] [PubMed] [Google Scholar]
  7. Dretzen G., Bellard M., Sassone-Corsi P., Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981 Apr;112(2):295–298. doi: 10.1016/0003-2697(81)90296-7. [DOI] [PubMed] [Google Scholar]
  8. Galibert F., Chen T. N., Mandart E. Nucleotide sequence of a cloned woodchuck hepatitis virus genome: comparison with the hepatitis B virus sequence. J Virol. 1982 Jan;41(1):51–65. doi: 10.1128/jvi.41.1.51-65.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Galibert F., Mandart E., Fitoussi F., Tiollais P., Charnay P. Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli. Nature. 1979 Oct 25;281(5733):646–650. doi: 10.1038/281646a0. [DOI] [PubMed] [Google Scholar]
  10. Hofstra H., Witholt B. Heat-labile enterotoxin in Escherichia coli. Kinetics of association of subunits into periplasmic holotoxin. J Biol Chem. 1985 Dec 15;260(29):16037–16044. [PubMed] [Google Scholar]
  11. Itoh Y., Takai E., Ohnuma H., Kitajima K., Tsuda F., Machida A., Mishiro S., Nakamura T., Miyakawa Y., Mayumi M. A synthetic peptide vaccine involving the product of the pre-S(2) region of hepatitis B virus DNA: protective efficacy in chimpanzees. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9174–9178. doi: 10.1073/pnas.83.23.9174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lederberg E. M., Cohen S. N. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. doi: 10.1128/jb.119.3.1072-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Milich D. R. Genetic and molecular basis for T- and B-cell recognition of hepatitis B viral antigens. Immunol Rev. 1987 Oct;99:71–103. doi: 10.1111/j.1600-065x.1987.tb01173.x. [DOI] [PubMed] [Google Scholar]
  15. Netter H. J., Guldner H. H., Szostecki C., Lakomek H. J., Will H. A recombinant autoantigen derived from the human (U1) small nuclear RNP-specific 68-kd protein. Expression in Escherichia coli and serodiagnostic application. Arthritis Rheum. 1988 May;31(5):616–622. doi: 10.1002/art.1780310506. [DOI] [PubMed] [Google Scholar]
  16. Neurath A. R., Kent S. B., Parker K., Prince A. M., Strick N., Brotman B., Sproul P. Antibodies to a synthetic peptide from the preS 120-145 region of the hepatitis B virus envelope are virus neutralizing. Vaccine. 1986 Mar;4(1):35–37. doi: 10.1016/s0264-410x(86)80001-9. [DOI] [PubMed] [Google Scholar]
  17. Okamoto H., Imai M., Usuda S., Tanaka E., Tachibana K., Mishiro S., Machida A., Nakamura T., Miyakawa Y., Mayumi M. Hemagglutination assay of polypeptide coded by the pre-S region of hepatitis B virus DNA with monoclonal antibody: correlation of pre-S polypeptide with the receptor for polymerized human serum albumin in serums containing hepatitis B antigens. J Immunol. 1985 Feb;134(2):1212–1216. [PubMed] [Google Scholar]
  18. Pasek M., Goto T., Gilbert W., Zink B., Schaller H., MacKay P., Leadbetter G., Murray K. Hepatitis B virus genes and their expression in E. coli. Nature. 1979 Dec 6;282(5739):575–579. doi: 10.1038/282575a0. [DOI] [PubMed] [Google Scholar]
  19. Sadoff J. C., Ballou W. R., Baron L. S., Majarian W. R., Brey R. N., Hockmeyer W. T., Young J. F., Cryz S. J., Ou J., Lowell G. H. Oral Salmonella typhimurium vaccine expressing circumsporozoite protein protects against malaria. Science. 1988 Apr 15;240(4850):336–338. doi: 10.1126/science.3281260. [DOI] [PubMed] [Google Scholar]
  20. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smith B. P., Reina-Guerra M., Stocker B. A., Hoiseth S. K., Johnson E. Aromatic-dependent Salmonella dublin as a parenteral modified live vaccine for calves. Am J Vet Res. 1984 Nov;45(11):2231–2235. [PubMed] [Google Scholar]
  22. Svennerholm A. M., Wikström M., Lindblad M., Holmgren J. Monoclonal antibodies to Escherichia coli heat-labile enterotoxins: neutralising activity and differentiation of human and porcine LTs and cholera toxin. Med Biol. 1986;64(1):23–30. [PubMed] [Google Scholar]
  23. Weimer T., Salfeld J., Will H. Expression of the hepatitis B virus core gene in vitro and in vivo. J Virol. 1987 Oct;61(10):3109–3113. doi: 10.1128/jvi.61.10.3109-3113.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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