水稻转高赖氨酸蛋白质基因(sb401)植株的获得及种子中蛋白质和氨基酸的含量分析

Transfer of High Lysine Gene sb401 into Rice and Analysis for Protein and Amino Acid Content in Transgenic Rice Seeds

用来源于马铃薯花粉的高赖氨酸蛋白质基因和玉米醇溶蛋白基因启动子构建了一个植物表达载体。采用农杆菌介导法导入籼稻品种龙特甫B ,PCR和Southernblot结果表明外源基因已整合到水稻基因组中,Westernblot分析表明高赖氨酸蛋白质基因已在转基因水稻种子中表达。对7株转基因水稻种子的分析表明,蛋白质的含量(10 . 87%～13. 16 % )平均提高了18. 7% ,赖氨酸的含量(0 . 4 7%～0 . 6 0 % )平均提高了10 % ,苏氨酸的含量(0 . 4 2 %～0 . 5 3% )平均提高了15 . 71% ,苯丙氨酸、蛋氨酸、组氨酸、缬氨酸、异亮氨酸和亮氨酸等必需氨基酸的含量也均有不同程度的提高。

The expression vector was constructed with lysine-rich protein gene sb401 from potato pollen and the p romoter of maize prolamin gene. The cultivar LongTeFuB was transfected with the above expression vector by Agrobacterium tumefaciens. Genomic PCR, Sou t hern-blotting analysis showed that the foreign gene was integrated into the ric e genome. The Western-blotting analysis confirmed that the transfected protein gene conferred high lysine were expressed in the mature seeds of the transgenic plants. The analysis result of mature seeds from 7 transgenic plants revealed t h at the content of protein was 10.87%-13.16% with the increase more than 18.7 %, the content of lysine was 0.47%-0.60% with the increase more than 10%; the c on tent of threonine (0.42%-0.53%) increased by 15.71%; and the contents of phe nyl alanine, methionine, histidine, valine leucine and isoleucine were also incre ased respectively.