【摘要】 甘油-3-磷酸酰基转移酶(glycerol-3-phosphate acyltransferase, GPAT)在植物三酰甘油脂(TAGs)等油脂合成中起着重要作用,而GPAT9基因是GPAT基因家族中参与油脂合成的重要基因。本研究采用电子克隆技术,以甘蓝型油菜湘油15为模板克隆了BnGPAT9基因的启动子序列pBnGPAT9,长度为900 bp。经过PlantCARE在线分析表明:该序列含有CAAT-box和TATA-box等区域特征元件,和多个参与光应答、激素诱导响应、逆境胁迫响应的元件,以及分生组织表达相关元件CAT-box与胚乳表达所需的顺式作用元件Skn-1_motif等,推测其可能在种子中表达较强烈,且在油菜生殖生长中有重要作用。通过双酶切,将pBnGPAT9启动子序列替换p BI121植物表达载体上的CaMV35S,使该启动子与GUS基因融合,构建了pBnGPAT9:GUS植物表达载体。更多还原

【Abstract】 Glycerol-3-phosphate acyltransferase(GPAT) plays an important role in oil synthesis such as plant tri acylglycerols(TAGs), and GPAT9 gene is an important gene involved in lipid synthesis in the GPAT gene family. In this study, the promoter sequence p BnGPAT9 of BnGPAT9 gene was cloned from Brassica napus Xiangyou 15 using electron cloning technology and the length was 900 bp. PlantCARE online analysis showed that the sequence contained region-specific elements such as CAAT-box and TATA-box, and multiple elements involved in photoresponse, hormone-induced response, and stress response, and cis-acting elements Skn-1_motif required for the expression of CAT-box and endosperm in meristem. It was speculated that the sequence might be strongly expressed in seeds and might play an important role in the reproductive growth of rapeseed. By double enzyme digestion,pBnGPAT9 promoter sequence was substituted for CaMV35 S on pBI121 plant expression vector, and the promoter was fused with GUS gene to construct pBnGPAT9: GUS plant expression vector.更多还原