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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。
Wheat glutenin 5 subunits directly affect the baking quality of bread, but most of our wheat proteins lack this subunit. The full-length nucleotide sequence (2 75 0 bp) and the wheat glutenin 5 subunit gene promoter (Psub5) of the wheat glutenin 5 subunit gene were obtained from the Cheyenne by primer design and PCR amplification Nucleotide sequence (6 30 bp). The sequencing results showed that Psub5, a promoter specific for wheat grain, and sub5, a structural gene for improving wheat quality, were obtained. Based on the construction of 6 intermediate vectors, the expression vector pCAMBIA1 3 0 1 Psub5 sub5 nos containing the gene of interest was finally obtained by selecting and changing the corresponding restriction sites. Digestion electrophoresis and PCR identification showed that the sub5 expression vector was successfully synthesized. It is hoped that this expression vector will be used for the quality improvement of wheat through genetic engineering.