摘要
为探索DREB2ACA基因(constitutive activate DREB2A,组成型激活DREB2A基因)在植物抗盐反应中的功能及其在植物抗逆基因工程中的运用,用SOE-PCR(overlapping extension-PCR,折叠延伸PCR)方法克隆了DREB2ACA,并将其置于盐诱导的rd29A启动子控制下,构建了植物表达载体,并通过农杆菌介导的方法将目的基因转入烟草。Southern blot证明了目的基因成功整合到烟草基因组中。RT-PCR表明,目的基因受盐胁迫诱导表达。在盐处理条件下,转基因烟草比对照具有更好的生长状态,含有较高的光和色素,具有较高的光合速率及较低的MDA含量。说明DREB2ACA的表达提高了植物抗逆能力,该基因在抗盐领域具有广阔的应用前景。
In order to investigate the function of DREB2ACA in plant salt-tolerance reaction and thus to explore its application value in genetic engineering. DREB2ACA was cloned using SOE-PCR, and was constructed in plant expression vector driven by rd29A promoter. Tobacco plants were genetically transformed with the vector. Integration of the T-DNA was confirmed using southern blot and expression of the target gene under salt stress were tested using RT-PCR. Compared to the control, transgenic lines had higher photosynthesis pigments, enhanced photosynthesis rate and lower MDA level under salt condition. All results demonstrate that DREB2ACA is probably a promising candidate gene for drought improvement in crops.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2012年第11期42-48,共7页
China Biotechnology
基金
国家自然科学基金资助项目(31271419
31271793)
关键词
DREB2A
转基因
烟草
抗盐
DREB2A Genetic transformation Tobacco Sah tolerance
