Pirdel Leila, Hosseini Ahmad Zavaran, Kazemi Bahram, Rasouli Manoochehr, Bandehpour Mojgan, Soudi Sara
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Avicenna J Med Biotechnol. 2012 Oct;4(4):186-92.
Various prokaryotic and eukaryotic expression systems have been developed for the production of recombinant proteins. In the present study, we used a new protein expression system based on the Iranian Lizard Leishmania, a trypanosomatid protozoan as a host, for the expression of LPG3 gene from Leishmania infantum (L.infantum).
The LPG3 gene was cloned in the expression cassette for integration into the small subunit of the ribosomal RNA locus of Lizard Leishmania genome by electroporation. Expression of the recombinant LPG3 protein was confirmed by western blotting and immunofluorescence staining.
Western blotting confirmed the expression and production of rLPG3 protein. Immunofluoresence analysis also revealed the staining throughout the cytoplasm of transfected parasites, indicating that the protein has been expressed.
These results demonstrate that Leishmania cells can be suggested an expression system for the production of recombinant LPG3 (rLPG3) to further research in vaccine designing against leishmaniasis.
为生产重组蛋白，已开发出多种原核和真核表达系统。在本研究中，我们使用了一种基于伊朗蜥蜴利什曼原虫（一种锥虫类原生动物）的新型蛋白表达系统作为宿主，来表达婴儿利什曼原虫（L.infantum）的LPG3基因。
通过电穿孔将LPG3基因克隆到表达盒中，以整合到蜥蜴利什曼原虫基因组核糖体RNA基因座的小亚基中。通过蛋白质印迹法和免疫荧光染色确认重组LPG3蛋白的表达。
蛋白质印迹法证实了rLPG3蛋白的表达和产生。免疫荧光分析还显示转染寄生虫的整个细胞质都有染色，表明该蛋白已被表达。
这些结果表明，利什曼原虫细胞可作为生产重组LPG3（rLPG3）的表达系统，用于进一步研究抗利什曼病疫苗的设计。
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