摘要
为通过基因工程改善植物蛋白氨基酸组成,在获得种子特异性表达拟南芥蛋氨酸合成途径关键基因At D-CGS转基因大豆的基础上,通过自交繁殖和性状鉴定,筛选出纯合转基因株系中作CGS-ZG11,并利用该株系与普通大豆品种石豆5号杂交,选育出中作CGS14J890等含有At D-CGS基因、综合性状优良的大豆新品系。本研究通过PCR和Southern blot检测,证明目的基因At D-CGS整合到中作CGS-ZG11基因组,以单拷贝形式存在。RT-PCR、Western blot结果显示,At D-CGS在转录和翻译水平上稳定表达。高效液相色谱测定结果表明,中作CGS-ZG11籽粒蛋氨酸含量显著高于野生型,且含量在T6、T7和T8连续3代相对稳定。大豆新品系中作CGS14J890等蛋氨酸含量高、综合性状好,证明At D-CGS基因可在不同的遗传背景下稳定遗传与表达。
Soybean is one of the most important sources of plant protein,but the content of methionine（ Met）in soybean protein is inadequate,which limits its value. Recent researches showed that improving the plant protein and amino acids composition of the plant protein via bio- technology method is a promising strategy. In the previous study,we transformed At D- CGS（ which is a key gene in Met biosynthesis and is expressed specifically in seeds） into soybean. A homozygous At D- CGS transgenic soybean line,Zhongzuo CGS- ZG11,has been obtained by self- fertilization and phenotypic identification through many generations. Zhongzuo CGS- ZG11 was crossed with a conventional soybean variety of Shidou 5 which has good comprehensive characteristics,and generated progeny lines,such as Zhongzuo CGS14J890. The PCR and Southern blot confirmed that At D- CGS gene was inherited from T6 to T8generations of Zhongzuo CGS- ZG11. The RT- PCR and Western blot showed the expression of At D- CGS was stable in Zhongzuo CGS- ZG11. The Met content in different generations of Zhongzuo CGS- ZG11 was investigated by High Performance Liquid Chromatography（ HPLC）. Compared to the wild- type seeds,the content of total Met in the transgenic soybean seeds increased remarkably,and remained consistently high through generations. The high methionine content in Zhongzuo CGS14J890 further proved that At D- CGS could function consistently among different genetic background and generations.
出处
《中国油料作物学报》
CAS
CSCD
北大核心
2015年第6期789-796,共8页
Chinese Journal of Oil Crop Sciences
基金
国家转基因生物新品种培育重大科技专项(2015ZX08011-003)
农业部财政部现代农业产业技术体系建设专项(CARS-04)
中国农业科学院科技创新工程
