大豆科学2011，Vol.30Issue(2)：190-193,4.
拟南芥AtLACS9基因的克隆及其植物表达载体构建
Cloning and Plant Expression Vector Construction of AtLACS9 Gene from Arabidopsis Thaliana
摘要
Abstract
Triacylglycerol (TAG) is the major form of storage lipid in oilseeds, and acyl-coenzyme A (CoA) and glycerol-3-phosphate are precursors for the synthesis of TAG. The long-chain acyl-CoA synthetases (LACSs) catalyze the synthesis of acyl-CoA molecules and therefore influence TAG content in oilseeds. In this paper, the soybean seed-specific lectin promoter (lec) , which was digested with restriction enzymes from vector pBl-lec, was inserted into the multiple cloning site of the plant expression vector pCAMBIA 3301 (p3301) , and then the complete coding region cDNA of Arabidopsis thaliana LACS9 (AtLACS9) gene was amplified by reverse transcription PCR( RT-PCR) and subcloned into the reconstructed vector p3301 downstream of the lee promoter. This work aimed at laying a foundation for future transformation of soybean with the gene and then obtaining transgenic soybean with enhanced oil content. The results showed that the vector p3301 was successully reconstructed by inserting the lec promoter and was named p3301-lec; AtLACS9 had an open reading frame of 2076 bp and the corresponding protein consists of 691 amino acids; the seed-specific expression vector of AtLACS9, namely p3301-lec-AtLACS9,was constructed successfully and could be used in the next experiments.关键词
长链脂酰辅酶A合成酶/基因克隆/载体构建Key words
Long-chain acyl-CoA synthetases (LACSs)/Gene cloning/Vector construction分类
农业科技引用本文复制引用
赵欢欢,吴兴,张锋,李宏伟,王茅雁..拟南芥AtLACS9基因的克隆及其植物表达载体构建[J].大豆科学,2011,30(2):190-193,4.基金项目
转基因生物新品种培育科技重大专项资助项目(2009ZX08004-009B). （2009ZX08004-009B）
