Recombinant expression of the rhoptry protein Clag3.1 as a green fluorescent chimeric protein in Plasmodium falciparum | ||||
| Delta Journal of Science | ||||
| Article 9, Volume 36, Issue 3, December 2013, Page 201-211 PDF (1.56 MB) | ||||
| Document Type: Research and Reference | ||||
| DOI: 10.21608/djs.2013.139775 | ||||
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| Author | ||||
| Ahmed M. Ghoneim* | ||||
| Zoology Department, Faculty of Science, Damietta University, New Damietta, Egypt. | ||||
| Abstract | ||||
| Like all members of Apicomplexa, the motile form of Plasmodium, the merozoite, possesses an elaborate apical complex composed of 3 organelles; micronemes, rhoptries and dense granules. Invasion of erythrocytes by Plasmodium merozoites is a tightly regulated multi-step process that begins with the cooperation of proteins released from micronemes and rhoptries to mediate attachment of parasites to the host cell membrane and to establish the parasitophorous vacuole. RhopH complex is a very abundant rhoptry secretion composed of 3 protein members; RhopH1, RhopH2 and RhopH3. RhopH1 is represented by either Clag2, Clag3.1, or Clag9. RhopH2 was found to traffic to the rhoptries via its first 24 amino acids. To further address how the other members of RhopH complex proteins are trafficked to rhoptries, the present study established a new transgenic Plasmodium falciparum line expressing a large chimeric protein containing almost ¾ of Clag3.1 fused to the N-terminus of GFP via a linker region under the control of rhoph2 promoter and its signal peptide encoding sequence. The data showed that this chimera was expressed efficiently indicating the strength of rhoph2 promoter, however, the chimera was trapped in the early secretory pathway and did not reach rhoptries. Provided a correct folding of this protein, these data could imply that the middle part of Clag3.1 (aa 484-1064) contain some information that interfere with the targeting contained in the first 24 amino acids of RhopH2. The C-terminus of Clag3.1 could contain Clag3.1-specific rhoptry targeting information. Based on literature available in the international databases, this is the first report to successfully express such a large rhoptry chimeric GFP protein. | ||||
| Keywords | ||||
| words: green fluorescent protein; Clag3.1; chimeric proteins; targeting; rhoptry; Plasmodium | ||||
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