[1]钱丹丹,龚德顺,焦丽,等.大豆异黄酮合成关键酶基因的克隆及表达分析[J].大豆科学,2011,30(05):743-748.[doi:10.11861/j.issn.1000-9841.2011.05.0743]
　QIAN Dan-dan,GONG De-shun,JIAO Li,et al.Cloning and Expression Analysis of Key Enzymes Genes in Biosynthesis of Soybean Isoflavones[J].Soybean Science,2011,30(05):743-748.[doi:10.11861/j.issn.1000-9841.2011.05.0743]
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大豆异黄酮合成关键酶基因的克隆及表达分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第30卷 期数: 2011年05期 页码: 743-748 栏目: 出版日期: 2011-10-25
Title:
Cloning and Expression Analysis of Key Enzymes Genes in Biosynthesis of Soybean Isoflavones
文章编号:
1000-9841（2011）05-0743-06
作者:
钱丹丹1; 龚德顺2; 焦丽1; 翟莹1; 张海军1; 李景文1; 王英1; 王庆钰1
1.吉林大学 植物科学学院，植物种质资源与利用实验室，吉林 长春130062；
2.中国科学技术大学生命科学学院，合肥微尺度物质科学国家实验室，安徽 合肥 230027
Author(s):
QIAN Dan-dan1; GONG De-shun2; JIAO Li1; ZHAI Ying1; ZHANG Hai-jun1; LI Jing-wen1; WANG Ying1; WANG Qing-yu1
1.Plant Germplasm and Application Laboratory,Plant Science College,Jilin University,Changchun 130062,Jilin;?
2.Hefei National Laboratory for Physical Sciences at Microscale,School of Life Science,University of Science and Technology of China,Hefei 230026,Anhui,China
关键词:
大豆异黄酮; 查尔酮合酶; 查尔酮异构酶; 表达分析
Keywords:
Soybean isoflavones; Chalcone synthase; Chalcone Isomerase; Expression Analysis
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2011.05.0743
文献标志码:
A
摘要:
采用RT-PCR法克隆了大豆异黄酮生物合成途径中查尔酮合酶(Chalcone synthase，CHS)和查尔酮异构酶(Chalcone Isomerase，CHI)2个关键酶基因，并利用SOE-PCR技术成功将2个基因融合，将其均构建成原核表达载体，在大肠杆菌中进行了初步的表达研究，为进一步研究CHS和CHI 2种关键酶的功能奠定了基础。同时构建了真核表达载体，采用农杆菌转化法侵染大豆子叶节，获得了转基因抗性植株。
Abstract:
Chalcone Synthase(CHS)and Chalcone Isomerase(CHI)are two key enzymes in soybean isoflavones biosynthesis pathway,the cDNAs of CHS and CHI were cloned using RT-PCR method and these two genes were fused using SOE-PCR，then constructed E. coli expression vectors to study the expression.The eukaryocyte expression vectors were also constructed and transferred into soybean cotyledonary nodes using Agrobacterium-mediated transformation,and then obtained the resistant transgenic plants.

参考文献/References:

[1]刘春龙,李忠秋,孙海霞,等.大豆异黄酮的生理作用及其在医学方面的研究进展[J].大豆科学,2008,27(4)：693-696.(Liu C L,Li Z Q,Sun H X,et al.Research progress and physiological function of soybean isoflavone on medicine[J].Soybean Science,2008,27(4):693-696.)

[2]夏循礼,杨广笑,何光源.大豆异黄酮生物合成串联基因簇及其表达载体的构建[J].大豆科学2009,28(1):7-10.(Xia X L,Yang G X,He G Y.Design of tandem genes cluster and expression vectors for soybean isoflavone biosynthesis[J].Soybean Science,2009,28(1):7-10.)

[3]Shih C H,Chen Y,Wang M,et al.Accumulation of isoflavone genistin in transgenic tomato plants overexpressing a soybean isoflavone synthase gene[J].Journal of Agricultural and Food Chemistry,2008,56(14):5655-5661.

[4]Zhang H C,Liu J M,Lu H Y,et al.Enhanced flavonoid production in hairy root cultures of Glycyrrhiza uralensis Fisch by combining the over-expression of chalcone isomerase gene with the elicitation treatment[J].Plant Cell Reports,2009,28(8):1205-1213.

[5]Cain C C,Saslowsky D E,Walker R A,et al.Expression of chalcone synthase and chalcone isomerase proteins in Arabidopsis seedlings[J].Plant Molecular Biology,1997,35:377-381.

[6]Li G L,Qiao Y K,Yang S H.Study of the Agrobacterium-mediated transformation systems of soybean cotyledonary node[J].Acta Agronomica Sinica,2005,31(2):170-176.

[7]李艳萍,郎志宏,李敏,等.融合基因在转基因植物中的应用[J].中国生物工程杂志,2007,27(5):137-141.(Li Y P,Lang Z H,Li M,et al.The application of polyproteinin transgenic plants[J].China Biotechnology,2007,27(5):137-141.)

[8]Abdelhak E A,Barakate A,Barak M A.Coordinate expression and independent subcellular targeting of multiple proteins from a single transgene[J].Plant Physiology,2004,135:16-24.

[9]Felipe P D,Luke G A,Hues L E.Eunum pluribus:multipie proteins from a self-procesing polyprotein[J].Trends in Biotechnology,2006,24(2):68-75.

[10]Francois I E,Broekaert W F,Cammue B P.Different approaches for multitransgene-stacking in plants[J]．Plant Science,2002,163:281-295.

[11]孙鹤.多基因融合转化方法在转基因玉米中的研究[D].中国农业科学院，2009.(Sun H.The research of the multigene fusion transformation in transgenic maize[D].The Chinese Academy of Agricultural Sciences，2009.)

[12]Liu M,Yang J,Cheng Y Q,et al.Optimization of soybean(Glycine max?L.Merrill)in planta ovary transformation using a linear minimal GUS?gene cassette[J].Journal of Zhejiang University-Science B,2009,10(12):870-876.

[13]Zhu J H,Quan J L,He Y K,et al.Studies on the transformation of plasmid Prok Ⅱ into Agrobacterium tumefaciens?LBA4404 and C58C1[J].Journal of Northwest Sci-Tech University of Agriculture and Forestry(Nature Science Edition),2006,34(7):91-95.

[14]Di R,Purcell V,Collins G B.Production of transgenic soybean lines expressing the bean pod mottle virus coat protein precursor gene[J].Plant Cell Reports,1996,15:746-750.

[15]Yi J,Derynck M R,Chen L,et al.Differential expression of CHS7 and CHS8 genes in soybean[J].Planta,2010,231:741-753.

[16]Mckenize K S,Rutger J N.Genetic analysis of amylase content,alkali spreading score and grain dimensions in rice[J].Crop Science,1983,23(2):306-3l3.


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备注/Memo

基金项目：转基因生物新品种培育重大专项资助项目（2008ZX08004-003）；吉林省科技厅重点资助项目(20080204)；国家自然科学基金资助项目（30971808）。

第一作者简介：钱丹丹(1986-)，女，在读硕士，研究方向为作物遗传育种。E-mail:Qiandandan2008K@163.com。
通讯作者：王庆钰(1963-)，女，教授，主要从事作物遗传育种研究。E-mail:wqy414cn@yahoo.com.cn。

更新日期/Last Update: 2014-08-15