Kourtz Lauralynn, Dillon Kevin, Daughtry Sean, Peoples Oliver P, Snell Kristi D
Metabolix, Inc., 21 Erie St., Cambridge, MA 02139, USA.
Transgenic Res. 2007 Dec;16(6):759-69. doi: 10.1007/s11248-007-9067-1. Epub 2007 Feb 6.
Arabidopsis plants were transformed with a multi-gene construct for expression of the polyhydroxybutyrate (PHB) biosynthetic pathway containing a gene switch that can be activated by commercially available non-steroidal ecdysone analogs approved for use on some crops as pesticides. T(1) progeny of transgenic Arabidopsis plants were isolated and screened for PHB production in the presence of ecdysone analogs. T(2) progeny derived from selected T(1) lines were subjected to further analysis by comparing PHB production levels prior to treatment with inducing agent and 21 days after initiation of induction. Significant PHB production was delayed in many of the engineered plants until after induction. PHB levels of up to 14.3% PHB per unit dry weight were observed in young leaves harvested from engineered T(2) plants after applications of the commercial ecdysone analog Mimic. PHB in older leaves reached levels of up to 7% PHB per unit dry weight. This study represents a first step towards engineering a chemically inducible gene switch for PHB production in plants using inducing agents that are approved for field use.
用一个多基因构建体转化拟南芥植株,用于表达聚羟基丁酸酯(PHB)生物合成途径,该构建体包含一个基因开关,可被批准用于某些作物作为杀虫剂的市售非甾体蜕皮激素类似物激活。分离转基因拟南芥植株的T(1)代子代,并在蜕皮激素类似物存在的情况下筛选PHB的产生。通过比较诱导剂处理前和诱导开始后21天的PHB产生水平,对来自选定T(1)系的T(2)代子代进行进一步分析。许多工程植株中显著的PHB产生延迟到诱导后。在施用商业蜕皮激素类似物Mimic后,从工程T(2)植株收获的幼叶中观察到PHB水平高达每单位干重14.3%。老叶中的PHB达到每单位干重高达7%的水平。这项研究代表了朝着利用批准用于田间的诱导剂在植物中工程化化学诱导基因开关以生产PHB迈出的第一步。