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首页> 外文期刊>Applied Microbiology and Biotechnology >Towards a biotechnological platform for the production of human pro-angiogenic growth factors in the green alga Chlamydomonas reinhardtii
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Towards a biotechnological platform for the production of human pro-angiogenic growth factors in the green alga Chlamydomonas reinhardtii

机译：朝着生物技术平台，用于生产绿藻骨髓瘤中的人血管生成因子Reinhardtii

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The recent use of photosynthetic organisms such as Chlamydomonas reinhardtii in biomedical applications has demonstrated their potential for the treatment of acute and chronic tissue hypoxia. Moreover, transgenic microalgae have been suggested as an alternative in situ drug delivery system. In this study, we set out to identify the best available combination of strains and expression vectors to establish a robust platform for the expression of human pro-angiogenic growth factors, i.e., hVEGF-165, hPDGF-B, and hSDF-1, in biomedical settings. As a case study, combinations of two expression vectors (pOpt and pBC1) and two C. reinhardtii strains (UVM4 and UVM11) were compared with respect to hVEGF-165 transgene expression by determination of steady-state levels of transgenic transcripts and immunological detection of recombinant proteins produced and secreted by the generated strains. The results revealed the combination of the UVM11 strain with the pBC1 vector to be the most efficient one for high-level hVEGF-165 production. To assess the robustness of this finding, the selected combination was used to create hPDGF-B and hSDF-1 transgenic strains for optimized recombinant protein expression. Furthermore, biological activity and functionality of algal-produced recombinant pro-angiogenic growth factors were assessed by receptor phosphorylation and in vitro angiogenesis assays. The results obtained revealed a potentiating effect in the combinatorial application of transgenic strains expressing either of the three growth factors on endothelial cell tube formation ability, and thus support the idea of using transgenic algae expressing pro-angiogenic growth factors in wound healing approaches.
机译：最近在生物医学应用中使用如衣原体的光合生物（如衣原体）的生物体已经证明了它们治疗急性和慢性组织缺氧的潜力。此外，已经提出转基因微藻作为原位药物递送系统的替代品。在这项研究中，我们开始鉴定菌株和表达载体的最佳可用组合，以建立一种表达人血管生成生长因子，即HVEGF-165，HPDGF-B和HSDF-1的鲁棒平台生物医学设置。作为一种情况，通过测定转基因转录物和免疫学检测，将两种表达载体（POPT和PBC1）和两种C. Reinhardtii菌株（UVM4和UVM11）的组合进行比较。由所产生的菌株产生和分泌的重组蛋白质。结果表明，UVM11应变与PBC1载体的组合是高级HVEGF-165生产中最有效的。为了评估该发现的稳健性，所选组合用于产生HPDGF-B和HSDF-1转基因菌株以优化重组蛋白表达。此外，通过受体磷酸化和体外血管生成测定评估藻类产生的重组促血管生成因子的生物活性和功能。得到的结果揭示了在内皮细胞管形成能力中表达三种生长因子中的任一种的转基因菌株组合施用的增强效应，从而支持在伤口愈合方法中表达促血管生成增生因子的转基因藻类的想法。

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