Abstract:

CASase is thought to be the first key enzyme in β-ODAP biosynthetic pathways of Lathyrus sativus. 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.

Key words: L. sativus, CASase gene, Gateway, RNAi

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505 bp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pSGRNAi-CASase，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。

关键词: 山黧豆, &beta, -腈基丙氨酸合成酶基因, Gateway, RNAi载体构建