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盘基网柄菌作为恶性疟原虫环子孢子蛋白的表达宿主。

Dictyostelium discoideum as an expression host for the circumsporozoite protein of Plasmodium falciparum.

作者信息

Fasel N, Begdadi-Rais C, Bernard M, Bron C, Corradin G, Reymond C D

机构信息

Institute of Biochemistry, University of Lausanne, Epalinges, Switzerland.

出版信息

Gene. 1992 Feb 15;111(2):157-63. doi: 10.1016/0378-1119(92)90683-g.

Abstract

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.

摘要

我们利用细胞黏菌盘基网柄菌(Dd)来表达恶性疟原虫环子孢子蛋白(CS),它是一种抗疟疾亚单位疫苗的潜在成分。这是通过基于盘状菌素I编码基因启动子的表达载体实现的,在该载体中,我们将编码Dd前导肽的序列与几乎完整的CS编码区相连(pEDII-CS)。在简单的磷酸盐缓冲液中使细胞饥饿可诱导mRNA和蛋白质水平的CS产生。pH值或细胞密度的变化似乎不影响CS的合成。产生CS的细胞既可以在其正常底物细菌上生长,也可以在半合成培养基上生长,而不会影响CS的积累水平。根据其大小以及一组单克隆抗体对表位的识别判断,Dd中产生的CS似乎与天然寄生虫蛋白相似。我们构建了第二个表达载体,其中CS受Dd ras启动子的控制。然后可以通过外部添加cAMP来诱导CS积累。这样一个严格调控的启动子可能允许表达对细胞有潜在毒性的蛋白质。因此,Dd可能是一个有用的真核系统,用于生产重组蛋白,特别是来自恶性疟原虫等人类或动物寄生虫的重组蛋白。

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