@article{178e526488f547a3b319374afc3f7b2c,
title = "Assembly and expression of a synthetic gene encoding the antigen Pfs48/45 of the human malaria parasite Plasmodium falciparum in yeast",
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.",
keywords = "Pfs48/45, Plasmodium falciparum, Yeast",
author = "Milek, {Richard L.B.} and Stunnenberg, {Hendrik G.} and Konings, {Ruud N.H.}",
note = "Funding Information: This work was supported by the Dutch Ministry for Development Cooperation (DGIS/SO), contract number NL002701. This investigation also received financial support from the UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases and from the Commission of the European Community for Life Sciences and Technologies for Developing Countries. Special thanks are due to Brenda Pisa, Hans Visser, Ralf Triepels, and Annemieke Janssens for technical assistance in yeast culturing and analysis of recombinants, to the Unilever Research Laboratories for plasmid pUR2730 and for anti α-galactosidase serum, to Dr. Jung (Department of Organic Chemistry, University of T{\"u}bingen) for the Pfs48/45 specific peptides, to Dr. William Leenders for the P. pastoris clone expressing VPF 121 , and to Dr. Manja Boerman, Dr. Monique Slijper and Dr. Joan Betz for their critical comments.",
year = "2000",
month = feb,
day = "4",
doi = "10.1016/S0264-410X(99)00392-8",
language = "English",
volume = "18",
pages = "1402--1411",
journal = "Vaccine",
issn = "0264-410X",
publisher = "Elsevier BV",
number = "14",
}