摘要
β-腈基丙氨酸合成酶(CASase)是调控山黧豆(Lathyrus sativus)内源毒素β-ODAP合成的关键酶。本研究以山黧豆幼根RNA为模板,利用RT-PCR扩增了505bp的山黧豆CASase基因序列;通过Gateway BP反应将扩增片段连接到入门载体构建pENTR-CASase。经测序验证后,将目的片段插入到植物表达载体构建pSGRNAiCASase,并将其转化到农杆菌GV3101中;为进一步的遗传转化奠定了基础。
CASase is thought to be the first key enzyme in β-ODAP biosynthetic pathways of Lathyrus sati- vus. To investigate the function of CASase gene and breed novel lines with low β-ODAP and high sulfur- containing amino acids, a 505 bp fragment of CASase gene was cloned from radicle RNA of L. sativus through RT-PCR. And then, an entry clone vector pENTR-CASase was constructed through Gateway BP cloning. An RNAi transformation vector pSGRNAi-CASase was constructed. Finally, the RNAi vector was transformed into Agrobacterium tumefaciens strain GV3101, which laid a foundation for obtaining transgenic plants with CASase gene silenced.
作者
张明科
刘凤娟
陶英杰
胡鑫
李荣硕
徐全乐
ZHANG Ming-ke LIU Feng-juan TAO Ying-jie HU Xin LI Rong-shuo XU Quan-le(College Hospital, Northwest A＆F University, Yangling, Shaanxi Province 712100, China College of Life Sciences, Northwest Ag〉F University, Yangling, Shaanxi Province 712100, China)
出处
《草地学报》
CAS
CSCD
北大核心
2016年第5期1146-1149,共4页
Acta Agrestia Sinica
基金
中央高校基本科研业务费(2014YB040
2452015401)
国家自然科学基金(31401910)
中国博士后科学基金(2016M590975)
陕西省博士后科研项目(2016BSHEDZZ119)资助
