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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 isoflavone synthase gene overexpression transgenic soybean callus. METHODS: The T-DNA part of isoflavone synthase gene plasmid pCAMB IA1301-C IP was transferred into soybean genome and induced with transgenic rhizobium-mediated transgene method. Results The GUS staining method was used to confirm that the foreign plasmids were well expressed. The insertions of foreign gene fragments and IFS gene expression were confirmed by PCR and reverse transcription PCR (RT-PCR). Conclusion The introduction of exogenous isoflavone synthase gene into soybean genome and its expression using Rhizobium-mediated transgenic system can provide a basis for the development and utilization of soy isoflavones and for the biosynthesis and regulation of soybean isoflavones.