Hu H H, Jing C Q, Liu R, Li W D, Feng H G
College of Life Science and Technology, Sanquan School, Xinxiang Medical University, Xinxiang, China.
College of Life Science and Technology, Xinxiang Medical University, Xinxiang, China.
Genet Mol Res. 2015 Aug 10;14(3):9291-7. doi: 10.4238/2015.August.10.9.
The aim of this study was to clone the isoflavone synthase (IFS) gene and establish the recombinant Minshan Trifolium pratense. The IFS gene was cloned from the callus of Minshan T. pratense using reverse transcription-polymerase chain reaction. The plant expression vector pRI101-AN-IFS was constructed and introduced into Agrobacterium tumefaciens strain LBA4404, and then screened under cephalosporin. IFS expression was detected by reverse transcription-polymerase chain reaction. The IFS gene was cloned successfully. Sequence analysis indicated that IFS gene had high homology with similar genes from other plants. The IFS-overexpressing callus was obtained by introducing the LBA4404-harboring IFS-pRI101-AN-IFS vector into T. pratense calluses.
本研究的目的是克隆异黄酮合酶（IFS）基因并构建重组岷山红三叶。采用逆转录-聚合酶链反应从岷山红三叶愈伤组织中克隆IFS基因。构建植物表达载体pRI101-AN-IFS并导入根癌农杆菌LBA4404菌株，然后在头孢菌素筛选下进行筛选。通过逆转录-聚合酶链反应检测IFS表达。成功克隆了IFS基因。序列分析表明，IFS基因与其他植物的相似基因具有高度同源性。通过将携带IFS-pRI101-AN-IFS载体的LBA4404导入红三叶愈伤组织，获得了过表达IFS的愈伤组织。
