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Dictyostelium discoideum as an expression host for the circumsporozoite protein of Plasmodium falciparum.
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We have used the cellular slime mold, Dictyostelium discoideum (Dd), to express the Plasmodium falciparum circumsporozoite protein (CS), a potential component of a subunit vaccine against malaria. This was accomplished via an expression vector based on the discoidin I-encoding gene promoter, in which we linked a sequence coding for a Dd leader peptide to the almost complete CS coding region (pEDII-CS). CS production at both the mRNA and protein levels is induced by starving cells in a simple phosphate buffer. Variation in pH or cell density does not seem to influence CS synthesis. CS-producing cells can be grown either on their normal substrate, bacteria, or on a semi-synthetic media, without affecting CS accumulation level. The CS produced in Dd seems similar to the natural parasite protein as judged by its size and epitope recognition by a panel of monoclonal antibodies. We constructed a second expression vector in which the CS is under the control of a Dd ras promoter. CS accumulation can then be induced by external addition of cAMP. Such a tightly regulated promoter may allow expression of proteins potentially toxic to the cell. Thus, Dd could be a useful eukaryotic system to produce recombinant proteins, in particular from human or animal parasites like P. falciparum.
Amino Acid Sequence, Animals, Antigens, Protozoan/chemistry, Antigens, Protozoan/genetics, Blotting, Western, Cloning, Molecular, Dictyostelium/genetics, Fungal Proteins/genetics, Gene Expression Regulation/drug effects, Genes, ras/genetics, Lectins, Molecular Sequence Data, Plasmodium falciparum/genetics, Promoter Regions, Genetic/genetics, Protozoan Proteins/chemistry, Protozoan Proteins/genetics, Recombinant Proteins/chemistry, Recombinant Proteins/genetics
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