XIA Xun-li,YANG Guang-xiao,HE Guang-yuan.Design of Tandem Genes Cluster and Expression Vectors for Soybean Isoflavone Biosynthesis[J].Soybean Science,2009,28(01):7-10,20.[doi:10.11861/j.issn.1000-9841.2009.01.0007]
大豆异黄酮生物合成串联基因簇及其表达载体的构建
- Title:
- Design of Tandem Genes Cluster and Expression Vectors for Soybean Isoflavone Biosynthesis
- 文章编号:
- 1000-9841(2009)01-0007-04
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 植物异黄酮是一种重要的次级代谢产物,在植物的生理活动以及人类健康方面具有重要作用。植物异黄酮生物合成途径有三个调控酶:查尔酮合酶(Chalcone synthase,CHS),查尔酮异构酶(Chalcone Isomerase,CHI)和异黄酮合酶(Isoflavone synthase,IFS)。以大豆为材料,利用RT-PCR方法克隆到查尔酮合酶基因(CHS),查尔酮异构酶基因(CHI Ⅱ)和异黄酮合酶基因(IFS)的cDNA(Genebank登记号分别为EU526827,EU526829,EU526830)经过NCBI在线Blast比对,相似度均在98%以上,确认克隆的目标基因是正确的;将这三种基因顺序连接,构建成串联基因簇CHS- CHI ⅡIFS(命名为rIFS),并且构建了rIFS的原核表达载体,进行了初步的表达研究。为进一步研究植物异黄酮的生物合成以及进行异黄酮生物工程奠定了基础。
- Abstract:
- Plant isoflavones is a sort of important secondary metabolites with signification in plant physiological function and healthiness for human being.There are three regulating enzymes in plant isoflavones biosynthesis pathway:Chalcone synthase(CHS),Chalcone isomerase(CHI)and Isoflavone synthase(IFS).In this article,we cloned the cDNAs of CHS,CHIⅡ,and IFS using RT-PCR method;then assembled these three genes to make a tandem genes cluster-CHS-CHI Ⅱ-IFS(we named it as rIFS);finally we constructed the E coli.expression vector of pET-rIFS and researched its expression.All these made the groundwork for further research on soybean isoflavone biosynthesis and isoflavone engineering.
参考文献/References:
[1]黄芸,秦民坚,余国英.异黄酮类化合物在植物界的分布及药理作用[J].中国野生植物资源,2001,20(1):5-7.(Huang Y,Qin M J,Yu G Y.Distributing of isoflavones in plants and their pharmacologicals functions[J].Chinese Wild Plant Resourses,2001,20(1):5-7.)
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备注/Memo
基金项目:国家重点基础研究发展计划(973计划)前期专项资助项目(2005CCA1900)。