Dictyostelium discoideum as an expression host for the circumsporozoite protein of Plasmodium falciparum.
Data(s) |
1992
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Resumo |
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. |
Identificador |
https://serval.unil.ch/?id=serval:BIB_0645695131F1 isbn:0378-1119 (Print) pmid:1541397 doi:10.1016/0378-1119(92)90683-G isiid:A1992HJ34800002 |
Idioma(s) |
en |
Fonte |
Gene, vol. 111, no. 2, pp. 157-163 |
Palavras-Chave | #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 |
Tipo |
info:eu-repo/semantics/article article |