Production of polyhydroxybutyrate by polycistronic expression of bacterial genes in tobacco plastid
Production of polyhydroxybutyrate by polycistronic expression of bacterial genes in tobacco plastid
复制标题
DOI：
10.1093/pcp/pch139
复制
发表时间：
2004-09-01
影响因子：
4.9
通讯作者：
Nakashita, H
中科院分区：
文献类型：
--
作者：
Arai, Y;Shikanai, T;Nakashita, H
转基因技术用于提高和改善作物产量，其在植物化学资源生产中的应用一直在研究中。为了实现后一个目标，需要多基因转化来改善或改变植物代谢途径;然而，当通过植物核转化完成时，该过程是昂贵且耗时的。我们成功地在烟草植物的代谢工程中引入多个基因内的细菌样操纵子到质体基因组。利用由大观霉素抗性基因和三个细菌基因组成的多顺反子转化烟草质体，对它们的核糖体结合位点进行修饰后合成生物可降解聚酯聚[（R）-3-羟基丁酸酯]（PHB）。DNA和RNA分析证实了引入的基因插入到质体基因组中，并且它们的多顺反子表达。结果表明，转质体烟草叶片中积累了大量的PHB。导入的基因和PHB生产力是母系遗传的，避免了花粉扩散的遗传传播，并在种子后代中稳定地保持。尽管低的聚羟基丁酸生产力，这份报告表明，transplastomic技术的代谢工程的可行性。这种“植物发酵”系统可应用于各种化学商品和药物的植物生产。
Transgenic techniques are used to enhance and improve crop production, and their application to the production of chemical resources in plants has been under investigation. To achieve this latter goal, multiple-gene transformation is required to improve or change plant metabolic pathways; when accomplished by plant nuclear transformation, however, this procedure is costly and time consuming. We succeeded in the metabolic engineering of the tobacco plant by introducing multiple genes within a bacteria-like operon into a plastid genome. A tobacco plastid was transformed with a polycistron consisting of the spectinomycin resistance gene and three bacterial genes for the biosynthesis of the biodegradable polyester, poly[(R)-3-hydroxybutyrate] (PHB), after modification of their ribosome binding sites. DNA and RNA analysis confirmed the insertion of the introduced genes into the plastid genome and their polycistronic expression. As the result, the transplastomic tobacco accumulated PHB in its leaves. The introduced genes and the PHB productivity were maternally inherited, avoiding genetic spread by pollen diffusion, and were maintained stably in the seed progeny. Despite the low PHB productivity, this report demonstrates the feasibility of transplastomic technology for metabolic engineering. This "phyto-fermentation" system can be applied to plant production of various chemical commodities and pharmaceuticals.