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| 山黧豆CASase基因的克隆及RNAi载体的构建 | |
| 引用本文： | 张明科,刘凤娟,陶英杰,胡鑫,李荣硕,徐全乐.山黧豆CASase基因的克隆及RNAi载体的构建[J].草地学报,2016,24(5):1146-1149. |
| 作者姓名： | 张明科 刘凤娟 陶英杰 胡鑫 李荣硕 徐全乐 |
| 作者单位： | 1. 西北农林科技大学医院, 陕西 杨凌 712100; 2. 西北农林科技大学生命科学学院, 陕西 杨凌 712100 |
| 基金项目： | 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) |
| 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 |
| 收稿时间： | 2015-02-02 |
Cloning of CASase Gene from Lathyrus sativus and Construction of its RNAi Vector |
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| ZHANG Ming-ke,LIU Feng-juan,TAO Ying-jie,HU Xin,LI Rong-shuo,XU Quan-le.Cloning of CASase Gene from Lathyrus sativus and Construction of its RNAi Vector[J].Acta Agrestia Sinica,2016,24(5):1146-1149. | |
| Authors: | ZHANG Ming-ke LIU Feng-juan TAO Ying-jie HU Xin LI Rong-shuo XU Quan-le |
| Institution: | 1. College Hospital, Northwest A & F University, Yangling, Shaanxi Province 712100, China; 2. College of Life Sciences, Northwest A & F University, Yangling, Shaanxi Province 712100, China |
| 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 sulfurcontaining 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. |
| Keywords: | L sativus CASase gene Gateway RNAi |
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