Document Detail


Assembly and expression of a synthetic gene encoding the antigen Pfs48/45 of the human malaria parasite Plasmodium falciparum in yeast.
MedLine Citation:
PMID:  10618538     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Pfs48/45 is an important transmission-blocking vaccine candidate antigen of the human malaria parasite Plasmodium falciparum. This study was aimed at synthesis of recombinant Pfs48/45 containing conformation-constrained epitopes of the native antigen in yeast. Since in the yeast Saccharomyces cerevisiae induction of gene-expression led to prematurely terminated transcripts, an entirely synthetic gene of higher GC content was assembled. Replacement of the AT rich natural gene by the synthetic gene relieved the observed premature transcription termination. Nevertheless, recombinant protein expression could not be detected. In contrast, in the yeast Pichia pastoris low levels of recombinant Pfs48/45 were produced upon induction of synthetic gene expression. The recombinant protein was shown to be disulphide-bridge constrained, but was not recognised by transmission-blocking antibodies and did not induce transmission-blocking sera in mice.
Authors:
R L Milek; H G Stunnenberg; R N Konings
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Vaccine     Volume:  18     ISSN:  0264-410X     ISO Abbreviation:  Vaccine     Publication Date:  2000 Jan 
Date Detail:
Created Date:  2000-03-07     Completed Date:  2000-03-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8406899     Medline TA:  Vaccine     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  1402-11     Citation Subset:  IM    
Affiliation:
Department of Molecular Biology and Cell Biology, University of Nijmegen, Toernooiveld 1, 6525 ED Nijmegen, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Base Sequence
DNA, Protozoan / genetics
Genes, Synthetic
Humans
Malaria / parasitology
Membrane Glycoproteins / biosynthesis,  genetics*,  immunology,  isolation & purification
Molecular Sequence Data
Pichia / genetics,  metabolism
Plasmodium falciparum / genetics*
Protozoan Proteins / biosynthesis,  genetics*,  immunology,  isolation & purification
Saccharomyces cerevisiae / genetics,  metabolism
Transfection
Chemical
Reg. No./Substance:
0/DNA, Protozoan; 0/Membrane Glycoproteins; 0/Protozoan Proteins; 0/pfs48-45 protein, Plasmodium falciparum

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