SBIR Phase IB: Regulating Transgene Expression for Biosecure Microalgal Biofuel Production

这个小型企业创新研究第一阶段项目开发了一个系统，在利用转基因藻类的同时，在开放的跑道上生物安全地生产藻油。通过生产含有特定基因调控序列的转基因藻类，这项研究开发了只有在特定诱导化合物存在的情况下才有条件地表达必要的叶绿体编码基因的藻类菌株。因此，转基因藻类菌株只有在诱导剂存在的情况下才能存活(诱导剂将被添加到我们生产设施的池塘中)。一旦逃离生产设施，这些藻类将停止生产必要的目标蛋白，并无法在环境中繁殖。这一生物安全生产系统将允许在开放式池塘生产系统中使用转基因藻类，而不必担心影响周围系统。拟议研究的更广泛影响将是能够使用低成本的开放式池塘养殖系统来生产生物燃料，使用通过遗传操作的微藻来最大限度地生产石油和生物质。这将以生物安全的方式实现，方法是开发分子生物学策略，防止基因操纵的藻类菌株一旦从生产池塘中移走就不再繁殖。利用分子遗传学既增加石油产量，又确保转基因藻类的生物安全使用(无法在周围环境中复制)，可以利用藻类系统以成本效益高的方式生产生物燃料。

This Small Business Innovation Research Phase I project develops a system for the biosecure production of algal oils in open raceways while utilizing genetically modified algae. Through production of transgenic algae containing specific gene regulatory sequences, this research develops algal strains that conditionally express an essential chloroplast-encoded gene only in the presence of the specific inducer compound. Thus, transgenic algal strains would survive only in the presence of the inducer compound (which will be added to the ponds in our production facility). On escape from the production facility, these algae will stop production of the essential target protein and be unable to reproduce in the environment. This biosecure production system will allow the use of genetically modified algae in open pond production systems without fear of impacting the surrounding systems. The broader impact of the proposed research will be to enable the use of low cost, open-pond culturing systems for the production of biofuels using microalgae genetically manipulated for maximum oil and biomass production. This will be accomplished in a biosecure manner through the development of molecular biological strategies that prevent the genetically manipulated algal strains from reproducing once they are removed from the production ponds. The use of molecular genetics to both increase the production of oil and ensure biosecure use of the transgenic algae (unable to replicate in the surrounding environment) could enable cost effective production of biofuels using algal systems.