小麦高分子量麦谷蛋白5亚基基因的克隆及其表达载体的构建(英文)

Cloning of Wheat High Molecular Weight Glutenin Subunit 5 Gene and Construction of Its Expression Vector

小麦麦谷蛋白 5亚基直接影响面包的烘烤品质 ,但我国大部分小麦品种的蛋白中缺少这种亚基。通过引物设计、PCR扩增 ,从Cheyenne中得到了小麦麦谷蛋白 5亚基结构基因 (sub5 )的全长核苷酸序列 (2 75 0bp)和小麦麦谷蛋白 5亚基基因启动子 (Psub5)的核酸序列 (6 3 0bp)。测序结果表明 :得到了小麦籽粒中特异表达启动子———Psub5和用于改良小麦品质的结构基因———sub5。通过选择和改变相应的酶切位点 ,在构建 6个中间载体的基础上 ,最后得到了含有目的基因的表达载体pCAMBIA1 3 0 1 Psub5 sub5 nos。酶切电泳及PCR鉴定表明 :已成功地合成了sub5的表达载体。有希望通过基因工程的方法将该表达载体用于小麦的品质改良。

Using the PCR amplification technology, a full-length nucleotide sequence (2 750 bp) of wheat high molecular weight glutenin subunit 5 (or Dx5) gene(Glu-1D-1d) and its promoter sequence (630 bp), were cloned from 'Cheyenne' wheat. By the selection and reparation of corresponding sites of restriction enzymes, a chimaeric gene, comprising the promoter (P sub5), the structure gene(Glu-1D-1d) and the nopaline synthase (Nos) gene terminator, i.e. pCAMBIA1301-P HGBy9-sub5-nos, was successfully constructed and verified after five transitional plasmids being produced.