Article: article from journal or magazin.
Dictyostelium discoideum as an expression host for the circumsporozoite protein of Plasmodium falciparum.
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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