Hasunuma Tomohisa, Kondo Akihiko, Miyake Chikahiro
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Japan.
Methods Mol Biol. 2010;643:213-27. doi: 10.1007/978-1-60761-723-5_15.
Plants synthesize a large number of isoprenoid compounds that have diverse structures and functions. All isoprenoids are synthesized through consecutive condensation of five-carbon precursors, isopentenyl diphosphate (IPP) and its allyl isomer dimethylallyl diphosphate (DMAPP). With recent success in the cloning of genes that encode the enzymes of isoprenoid biosynthesis, genetic engineering strategies for the improvement of plant isoprenoid metabolism have emerged. Plastid transformation technology offers attractive features in plant genetic engineering. It has many advantages over nuclear genome transformation: high-level foreign protein expression, no need for a transit peptide, absence of gene silencing, and convenient transgene stacking in operons. We demonstrated that this technology is a remarkable tool for the production of isoprenoids in plants through metabolic engineering. The expression of bacterial genes encoding CrtW (beta-carotene ketolase) and CrtZ (beta-carotene hydroxylase) or cyanobacterial genes encoding DXR (1-deoxy-D-xylulose-5-phosphate reductoisomerase) in the plastid genome leads to alteration in isoprenoid content of tobacco leaves.
植物合成大量具有不同结构和功能的类异戊二烯化合物。所有类异戊二烯都是通过五碳前体异戊烯基二磷酸(IPP)及其烯丙基异构体二甲基烯丙基二磷酸(DMAPP)的连续缩合合成的。随着最近在克隆编码类异戊二烯生物合成酶的基因方面取得成功,出现了用于改善植物类异戊二烯代谢的基因工程策略。质体转化技术在植物基因工程中具有吸引人的特点。与核基因组转化相比,它有许多优点:高水平的外源蛋白表达、无需转运肽、不存在基因沉默以及便于在操纵子中进行转基因堆叠。我们证明,这项技术是通过代谢工程在植物中生产类异戊二烯的一项卓越工具。在质体基因组中表达编码CrtW(β-胡萝卜素酮酶)和CrtZ(β-胡萝卜素羟化酶)的细菌基因或编码DXR(1-脱氧-D-木酮糖-5-磷酸还原异构酶)的蓝细菌基因会导致烟草叶片类异戊二烯含量的改变。