摘要
[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。
[ 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.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2013年第2期300-306,共7页
Xinjiang Agricultural Sciences
基金
新疆维吾尔自治区自然科学基金项目(2010211A01)
新疆维吾尔自治区科技支疆项目(20091129)
新疆维吾尔自治区高校科研计划重点项目(XJEDU2008I0)
新疆生物资源基因工程重点实验室开放基金项目(XJDX0201-2009-03)
关键词
苦豆子凝集素基因
载体构建
烟草
转化
Sophora alopecuroides lectin gene
vector construction
tobacco
transformation
