[1]谢璐遥,李晓琳,罗利,等.大豆GmSOT1基因克隆、载体构建及初步表达研究[J].大豆科学,2014,33(06):815-819.[doi:10.11861/j.issn.1000-9841.2014.06.0815]
　XIE Lu-yao,LI Xiao-lin,LUO Li,et al.Cloning,Plant Expression Vector Construction and Preliminary Study of GmSOT1 Gene from Soybean[J].Soybean Science,2014,33(06):815-819.[doi:10.11861/j.issn.1000-9841.2014.06.0815]
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大豆GmSOT1基因克隆、载体构建及初步表达研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第33卷 期数: 2014年06期 页码: 815-819 栏目: 出版日期: 2014-12-25
Title:
Cloning,Plant Expression Vector Construction and Preliminary Study of GmSOT1 Gene from Soybean
文章编号:
1000-9841.2014.06.0815
作者:
谢璐遥1; 2; 李晓琳2; 罗利3; 文涛1; 朱木兰2
1.四川农业大学 农学院，四川 成都 611130；
2.中国科学院 上海生命科学研究院 植物生理生态研究所/国家植物基因研究中心（上海)，上海 200032;
3.上海大学 生命科学学院/上海市能源植物育种与应用重点实验室,上海 200444
Author(s):
XIE Lu-yao1; 2; LI Xiao-lin2; LUO Li3; WEN Tao1; ZHU Mu-lan2
1.Agricultural College,Sichuan Agricultural University,Chengdu 611130,China;
2.Institute of Plant Physiology & Ecology,Shanghai Institutes for Biological Sciences,CAS,National Center for Plant Gene Research(Shanghai),Shanghai 200032,China;
3.Shanhai Key Lab of Bioenergy Plant,School of Life Science,Shanghai University,Shanghai 200444,China
关键词:
大豆; 磺酸基转移酶; 硫代葡萄糖苷; 基因克隆; 载体构建
Keywords:
Soybean; Sulfotransferase(SOT); Glucosinolate; Gene cloning; Vector construction
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2014.06.0815
文献标志码:
A
摘要:
磺酸基转移酶（sulphotransferase，简称SOT)是硫代葡萄糖苷（glucosinolate，简称硫苷)核心结构合成途径中的关键酶之一。本研究采用反向遗传学方法从大豆基因组中克隆GmSOT1基因并对其功能进行初步研究。首先采用PCR方法扩增大豆-GmSOT1-基因，并将其连接到pRNAi载体上的35S（CaMV35S)启动子后面；将35S:GmSOT1-基因片段从pRNAi载体上双酶切下来，连接到植物双元表达载体pCAMBIA2301的多克隆位点上，且命名为pCAMBIA2301-35S:GmSOT1；采用根癌农杆菌介导的遗传转化方法转化大豆成熟胚，并考察其初步表达结果。结果表明：大豆-GmSOT1-基因的编码区（coding sequences，简称CDS)由1 014 bp的核苷酸组成，编码一个长度为337个氨基酸残基的蛋白质；35S:GmSOT1被成功导入大豆胚细胞，初步观察发现GmSOT1过量表达导致大豆胚根明显膨大弯曲。
Abstract:
Sulfotransferase(SOT)is one of the most important key enzymes involved in the biosynthesis of the core structure of glucosinolate.Here GmSOT1 was cloned from the soybean genome using reverse genetics to investigate its preliminary functions.An ORF(open reading frame)DNA of GmSOT1 was amplified by PCR and ligated into a pRNAi vector under control of the 35S(CaMV35S)promoter.The 35S:GmSOT1 DNA fragment was excised from the pRNAi vector and ligated into the multiple cloning sites of pCAMBIA2301,named as pCAMBIA2301-35S:GmSOT1.The 35S-GmSOT1 construct was introduced into mature embryo cells of soybean through Agrobacterium-mediated genetic transformation.The transformed soybean embryos were used for preliminary phenotype analysis.Our results showed that the cloned GmSOT1 gene(CDS)-consisted of 1 014 bp nucleotide acids and encoded a 337 amino-acid protein.The construct of 35S:GmSOT1 was successfully introduced into soybean embryo cells.The transformed radicles appear enlarged and curved.The results provide a new cue to further study on the function of GmSOT1.

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备注/Memo

基金项目：国家高技术研究发展计划“863计划”（2013AA102704）。

第一作者简介：谢璐遥(1989-)，女，在读硕士，主要从事大豆基因克隆、载体构建和遗传转化研究。E-mail：shieluyau@yeah.net。

更新日期/Last Update: 2014-12-25