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首页> 外文期刊>Chinese science bulletin >Expression of lysine-rich protein gene and analysis of lysine content in transgenic wheat
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Expression of lysine-rich protein gene and analysis of lysine content in transgenic wheat

机译：富含赖氨酸蛋白基因的表达及转基因小麦中赖氨酸含量的分析

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摘要

Expression vector pBPC102, which carries winged bean lysine-rich protein (wblrp) gene and dihydropi-colinate synthase (DHDPS) gene, was transferred into hexap-loid winter wheat cv. Jinghua No.l, Jing411, You899 and Yangnong 15 explants of immature inflorescence and immature embryos by particle bombardment. More than 100 transgenic plants were obtained under the selection of s-(2-aminoethyl)-L-cysteine (AEC). Confirmed transgenic plants of T_0 and T_1 generation by PCR and PCR-Southern blotting analyses showed successful integration of wblrp gene into wheat genome. Analysis of transgenic plant lines of T_2 by Northern dot-blotting showed good expression of wblrp gene in offspring seed. The content of free lysine in leaves, contents of bound lysine and total proteins in seeds of T_2 transgenic wheat lines were determined and analyzed. Among 34 tested transgenic lines, levels of free lysine content in leaves of 9 transgenic lines are 2~3times higher than un-trans-formed wild-type cultivars. Among 17 analyzed transgenic lines, bound lysine content of 4 transgenic lines is more than 10% higher than that of wild-type cultivars. Our research suggests that introducing wblrp gene into wheat is an effective way to improve its nutrition quality.
机译：携带有翼豆富含赖氨酸的蛋白质（wblrp）基因和二氢吡啶-亚麻酸合酶（DHDPS）基因的表达载体pBPC102被转移到六倍体冬小麦简历中。经粒子轰击的京华1号，京411号，油烟899号和养农15个未成熟花序和未成熟胚的外植体。通过选择s-（2-氨基乙基）-L-半胱氨酸（AEC）获得了100多种转基因植物。通过PCR和PCR-Southern印迹分析确认的T_0和T_1世代转基因植物显示wblrp基因成功整合到小麦基因组中。通过Northern斑点印迹法对T_2转基因植物株系的分析表明，wblrp基因在后代种子中表达良好。测定并分析了T_2转基因小麦品系种子中叶片中游离赖氨酸的含量，结合赖氨酸的含量以及总蛋白的含量。在34个测试的转基因品系中，9个转基因品系的叶片中游离赖氨酸含量水平是未转化的野生型品种的2〜3倍。在分析的17个转基因品系中，4个转基因品系的结合赖氨酸含量比野生型品种高10％以上。我们的研究表明，将wblrp基因导入小麦是提高其营养品质的有效途径。

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