作物学报 ›› 2016, Vol. 42 ›› Issue (11): 1629-1637.
石磊,苗利娟,齐飞艳,张忠信,高伟,孙子淇,黄冰艳,董文召,汤丰收,张新友*
SHI Lei, MIAO Li-Juan, QI Fei-Yan, ZHANG Zhong-Xin, GAO Wei, SUN Zi-Qi, HUANG Bing-Yan, DONG Wen-Zhao, TANG Feng-Shou, and ZHANG Xin-You*
摘要:
Δ9-硬脂酰-ACP脱氢酶(SAD)是决定植物体内饱和脂肪酸与不饱和脂肪酸比值的关键酶。以花生品种豫花9326基因组DNA为模板,通过基因组步移技术,克隆到花生Δ9-硬脂酰-ACP脱氢酶基因(AhSAD)起始密码子ATG上游720 bp片段,利用5? RACE方法获得了该基因的5? UTR序列,通过序列比对确定720 bp片段为AhSAD启动子区域。PLACE在线启动子预测分析表明,该序列具有真核生物启动子必需的核心元件TATA-box和CAAT-box,含有多个与光诱导和激素响应相关顺式序列元件。将AhSAD启动子片段替换pBI121质粒中的CaMV35S启动子驱动下游GUS基因表达,构建植物表达载体pBI-PAhSAD。通过农杆菌介导法转化拟南芥和在花生不同组织中瞬时表达,利用GUS组织化学染色研究其表达特性。表明,在拟南芥和花生受体中,AhSAD启动子主要调控下游基因在根、茎、叶片和子叶中表达,在花生的果针中也检测到GUS活性;拟南芥的茎生叶只有叶脉中具有GUS活性,而花生整个叶片中都具有GUS活性。
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