Expression of the core antigen gene of hepatitis B virus (HBV) in Acetobacter methanolicus using broad-host-range vectors

Using the broad-host-range promoter probe vector pRS201 for cloning of phage Acm1 promoters, we established a convenient vector system for expression of heterologous genes in different Gram-negative bacteria. The usefulness of this system was demonstrated by expression of the HBV core gene in Acetob...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 1991-08, Vol.35 (5), p.631-637
Main Authors: SCHRODER, R, MAASSEN, A, LIPPOLDT, A, BORNER, T, BON BAEHR, R, DOBROWOLSKI, P
Format: Article
Language:English
Subjects:
Acetobacter - genetics
Amino Acid Sequence
Base Sequence
Biological and medical sciences
Biotechnology
Cloning, Molecular - methods
DNA, Recombinant
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Genes, Viral
Genetic engineering
Genetic Markers
Genetic technics
Genetic Vectors
Hepatitis B Core Antigens - genetics
hepatitis B virus
Hepatitis B virus - genetics
Methods. Procedures. Technologies
Modification of gene expression level
Molecular Sequence Data
Oligodeoxyribonucleotides
Plasmids
Restriction Mapping
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Summary:Using the broad-host-range promoter probe vector pRS201 for cloning of phage Acm1 promoters, we established a convenient vector system for expression of heterologous genes in different Gram-negative bacteria. The usefulness of this system was demonstrated by expression of the HBV core gene in Acetobacter methanolicus. Plasmids carrying the HBV core gene downstream of different Acm1-phage promoters were transferred to A. methanolicus, a new potential host for recombinant DNA expression. Using enzyme immunoassay and immunoblot techniques, the amount and composition of core antigen produced in A. methanolicus were compared with that derived from Escherichia coli. The expression of immunoreactive core antigen in A. methanolicus exceeds by sevenfold that in E. coli using an expression system with tandemly arranged promoters. Morphological observations by electron microscopy show that the HBV core gene products isolated from both hosts are assembled into regular spherical particles with a diameter of about 28 nm that are comparable to original viral nucleocapsids.
ISSN:0175-7598
1432-0614
DOI:10.1007/BF00169628