Assembly and expression of a synthetic gene encoding the antigen Pfs48/45 of the human malaria parasite Plasmodium falciparum in yeast

Richard L.B. Milek, Hendrik G. Stunnenberg, Ruud N.H. Konings

Research output: Contribution to journalArticlepeer-review

40 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1402-1411
Number of pages10
JournalVaccine
Volume18
Issue number14
DOIs
Publication statusPublished - 4 Feb 2000
Externally publishedYes

Keywords

  • Pfs48/45
  • Plasmodium falciparum
  • Yeast

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