[1]李兴龙,霍建玲,庞添,等.HSSP基因植物表达载体构建及其对大豆的遗传转化[J].大豆科学,2016,35(01):171-175.[doi:10.11861/j.issn.1000-9841.2016.01.0171]
　LI Xing-long,HUO Jian-ling,PANG Tian,et al.High Efficient Plant Expression Vector with HSSP Gene and Transformation into Soybean[J].Soybean Science,2016,35(01):171-175.[doi:10.11861/j.issn.1000-9841.2016.01.0171]
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HSSP基因植物表达载体构建及其对大豆的遗传转化

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年01期 页码: 171-175 栏目: 出版日期: 2016-01-20
Title:
High Efficient Plant Expression Vector with HSSP Gene and Transformation into Soybean
作者:
李兴龙; 霍建玲; 庞添; 柏锡
东北农业大学 生命科学学院，黑龙江 哈尔滨 150036
Author(s):
LI Xing-long ; HUO Jian-ling; PANG Tian; BAI Xi
College of Life Science of Northeast Agricultural University，Harbin 150030，China
关键词:
大豆; 含硫氨基酸; 农杆菌介导遗传转化
Keywords:
Soybean;  Sulfur-containing amino acids;  Agrobacterium mediated transformation
DOI:
10.11861/j.issn.1000-9841.2016.01.0171
文献标志码:
A
摘要:
大豆中含硫氨基酸匮乏，限制了大豆的营养价值和商业价值。为了提高大豆的含硫氨基酸含量，利用人工设计合成的高含硫氨基酸贮藏蛋白基因(HSSP)转化大豆。以载体pTF101-1为骨架，构建植物表达载体pTFGS，利用农杆菌介导法转化大豆品种东农50，经PCR及试纸条检测，获得转基因大豆16株，基因的转化效率为1.94%。对转基因大豆株系GSDL5进行组织特异性分析，结果表明，HSSP基因在种子中超量表达，而在其它组织部位仅有微量表达。采用接头PCR方法对株系GSDL5基因组插入位点侧翼序列进行克隆，获得与大豆基因组序列匹配的436 bp 侧翼序列，HSSP基因插入2号染色体基因的非编码区。经研究，获得了遗传背景明确的，仅在大豆种子中超量表达HSSP的转基因株系GSDL5。
Abstract:
Lack of sulfur amino acids limited the nutritional value and commercial value of soybeans.In order to improve the sulfur amino acid content, increase nutritional value of soybeans, this study was designed and synthesized artificially high sulfur amino acid storage protein gene (HSSP) and transformed into soybeans. The plant expression vector pTFGS was constructed with efficient plant expression vector pTF101-1 skeleton,via Agrobacterium-mediated transformation of soybean varieties Dongnong 50, detected 16 transgenic soybeans by PCR and strip, conversion efficiency was 1.94%. Transgenic lines GSDL5 were tissue-specific analysis showed that, HSSP gene overexpressed in seeds, while only trace expression in other tissue sites PCR method using the connector lines GSDL5 genomic sequences flanking the insertion site for cloning, obtained with soybean genome sequence flanking sequences matching 436 bp,HSSP gene is inserted into a non-coding region of chromosome 2.This study got a clear genetic background, and only in soybean seeds overexpressing HSSP transgenic lines GSDL5.

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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（2014ZX08004-003-002）。

第一作者简介：李兴龙（1988-），男，硕士，主要从事植物学研究。E-mail：xl__li@yeah.net。
通讯作者：柏锡（1975-），男，副教授，主要从事植物分子生物学研究。E-mail: baixi@neau.edu.cn。

更新日期/Last Update: 2016-02-05