异黄酮合成酶基因高表达的大豆转基因愈伤组织的研究  

Transgenic soybean callus with high expression of IFS gene

作　　者：姜楠[1] 王东[1] 崔征[1] 郑英秀 

机构地区：[1]沈阳药科大学中韩分子生药学研究室,辽宁沈阳110016 [2]韩国东亚大学生命资源科学学院分子育种研究室,韩国釜山604-714

出　　处：《沈阳药科大学学报》2009年第1期63-68,共6页Journal of Shenyang Pharmaceutical University

摘　　要：目的获得异黄酮合成酶基因高表达的转基因大豆愈伤组织。方法利用根瘤菌介导的大豆转基因方法,将异黄酮合称酶基因质粒pCAMB IA1301-C IP的T-DNA部分转入大豆基因组中并诱导转基因愈伤。结果利用GUS染色法,确认外源质粒片段基本表达良好;利用PCR及反转录PCR(RT-PCR)确定外源基因片段的插入及IFS基因的表达情况良好。结论利用根瘤菌介导的转基因体系,可将外源异黄酮合成酶基因转入大豆基因组中并实现其表达,可为开发利用大豆异黄酮,以及为大豆异黄酮生物合成与调控研究提供依据。Objective To obtain transgenic soybean callus with high expression of IFS gene. Methods By applying the Agrobacterium-mediated soybean transformation method, transfer the T-DNA of the plasmid PCAMBIA1301-CIP which contains the IFS gene was transferred into soybean genome and induced callus. Results GUS assay analysis showed the expression of exogenous gene. The integration and expression of IFS were confirmed by PCR and RT-PCR. Conclusions The exogenous IFS can be integrated and highly expressed into the soybean genome by the Agrobacterium-mediated method, and the result may be used as a alternative way to take advantage of the soy-isoflavonoids pathway.

关 键 词：异黄酮合成酶基因 PCR RT-PCR 根瘤菌介导 大豆转基因 GUS染色法 

分 类 号：R915[医药卫生—微生物与生化药学]