拟南芥2A6基因正义、反义植物表达载体的构建

Construction sense and antisense plant expression vectors of 2A6 gene in Arabidosis thaliana

利用PCR技术从番茄基因组中扩增了长约1.1kb的E8基因启动子,构建了中间表达载体pCAMBI-AE8;利用RT-PCR方法从拟南芥中扩增了长约1 197bp的2A6基因全长cDNA编码区,将其定向插入pCAMBI-AE8,构建了正义植物表达载体pCAMBIAE8-2A6;同时扩增了长约500bp的cDNA片段,定向插入pCAMBI-AE8,构建了反义植物表达载体pCAMBIAE8-2A6anti.以此为基础,可以进一步研究2A6基因的差异表达对转基因拟南芥角果发育的影响,达到揭示2A6基因功能的目的.

The 1 100 bp 5＇ flanking sequence of E8 gene was amplified by Polymerase Chain Reaction（PCR） from leaf tissue of tomato.The cloned DNA of E8 gene was introduced into vector pCAMBIA2300 to construct intermediate vector pCAMBIAE8.The 1 197 bp sequence of 2A6 gene was amplification by RT-PCR from Arabidosis thaliana.The cloned DNA of 2A6 gene was introduced into vector pCAMBIAE8 in a sense orientation,giving expression vector pCAMBIAE8-2A6.For constructing antisense expression vector pCAMBIAE8-2A6anti,500 bp sequence of 2A6 gene was cloned and introduced into vector pCAMBIAE8 in a antisense orientation.Based on them,the function of 2A6 gene will be showed by the differential expression of 2A6 gene in the maturity of siliques in transgenic Arabidosis in further research.