苦豆子凝集素基因植物表达载体的构建及其对烟草的转化

目的]了解苦豆子凝集素基因(SAL)的功能,将该基因构建到植物表达载体并转化烟草,获得转基因植株.方法]利用RT-PCR技术,提取苦豆子总RNA进行反转录得到cDNA,通过PCR扩增得到苦豆子凝集素基因SAL,并将其克隆到植物表达载体pCAMBIA1301上,产生重组质粒pCAMBIA1301-SAL.采用农杆菌介导法将重组质粒转化烟草,并进行分析鉴定.结果]构建了植物表达载体pCAMBIA1301-SAL,转化烟草后经筛选获得76株转基因植株,经抗性筛选及PCR和RT-PCR鉴定,其中27株显示为阳性植株.结论]苦豆子凝集素基因已经在烟草中成功表达,为进一步研究验证转基因烟草的抗病效果奠定了基础.

Construction of Plant Expression Vector Carrying Sophora alopecuroides Lectin Gene and Its Transformation to Tobacco

【Objective】To understand the role of Sophora alopecuroide lectin(SAL),SAL gene was used to construct the plant expressing vector and transformed tobacco for obtaining transgenic tobacco.【Method】Sophora alopecuroide lectin gene(SAL) was amplified from total RNA from Sophora alopecuroide leaf by RT -PCR technology.The amplified fragment was cloned into plant expression vector pCAMBIA1301 to generate recombinant plasmid pCAMBIA1301- SAL.The vector was transformed into to tobacco by agrobacterium mediated method.Agrobacterium - mediated transient expression assay was evaluated.【Result】Plant expression vector pCAMBIA1301 -SAL was successfully constructed,and transformed into tobacco.Seventy - six resistance plants were obtained.PCR results showed that 27 plants were positive.【Conclusion】Sophora alopecuroide lectin gene(SAL) was successfully expressed in tobacco.All of this laid foundations for the further study of plant disease resistance.