Expression of lysine-rich protein gene and analysis of lysine content in transgenic wheat

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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.1, Jing411, You899 and Yangnong15 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 T0 and T1 generation by PCR and PCR-Southern blotting analyses showed successful integration of wblrp gene into wheat genome. Analysis of transgenic plant lines of T2 by Northern dot-blotting showed good expression of wblrp gene in offspring seed. The content of free lysine in leaves, con- tents of bound lysine and total proteins in seeds of T2 trans- genic 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 effec- tive way to improve its nutrition quality. Jinghua No.1, Jing411, You899 and Yangnong15 explants of immature inflorescence (wdlrp) gene and was digested into hexaploids 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 T0 and T1 generation by PCR and PCR-Southern blotting analyzes showed successful integration of wblrp gene into wheat genome. Analysis of transgenic plant lines of T2 by Northern dot-blotting showed good expression of wblrp gene in offspring seed. The content of free lysine in leaves, con- tents of bound lysine and total proteins in seeds of T2 trans- genic 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 cultivator ars. 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 effec tive way to improve its nutrition quality.
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