Cloning and expression of a candidate malarial epitope in bacille Calmette Guerin

The candidate malarial vaccine epitope, region II of Plasmodium falciparum circumsporozoite protein (RII-CSP), was cloned into Mycobacterium smegmatis and M. bovis BCG via a mycobacterial replicative plasmid pUS1762. This plasmid contained a gene for the M. leprae18 kDa protein to provide expression...

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Bibliographic Details
Published in:Biotechnology letters 1997-11, Vol.19 (11), p.1135-1137
Main Authors: NORAZMI, M. N, DALE, J. W
Format: Article
Language:English
Subjects:
Bacteria
Biological and medical sciences
Biotechnology
Cloning
Deoxyribonucleic acid
DNA
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Methods. Procedures. Technologies
Modification of gene expression level
Proteins
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Summary:The candidate malarial vaccine epitope, region II of Plasmodium falciparum circumsporozoite protein (RII-CSP), was cloned into Mycobacterium smegmatis and M. bovis BCG via a mycobacterial replicative plasmid pUS1762. This plasmid contained a gene for the M. leprae18 kDa protein to provide expression signals. Transformation was achieved by electroporation and selection for kanamycin resistance and verified by Southern hybridisation and sequencing. The transformation efficiency was in the order of 10 cfu/μg DNA for M. smegmatis and 10 cfu/μg DNA for M. bovis BCG. Western blotting using a polyclonal antibody specific for the RII-CSP and a monoclonal antibody specific for the M. leprae protein showed the expected 25 kDa band of the 18 kDa-RII-CSP fusion protein.[PUBLICATION ABSTRACT]
ISSN:0141-5492
1573-6776
DOI:10.1023/A:1018405013737