小麦HMW-GS12基因克隆及植物表达载体构建

Cloning of HMW-GS 12 Gene in Wheat and Construction of Plant Expression Vectors

目的:为了利用基因遗传转化改良小麦品质,采用聚合酶链式反应(PCR)技术。方法:从小麦品种东农7742基因组DNA中扩增并克隆了小麦高分子量谷蛋白12亚基基因(HMW-GS 12)。结果:序列分析结果表明,该基因全长1 980bp,其核苷酸顺序和推导的氨基酸顺序与已发表的序列相比,同源性分别为99.5%和99.7%。经过基因拼接,分别构建了胚乳特异性表达和组成型表达的高分子量谷蛋白12亚基基因的两个植物表达载体pDNPPBIHG和pUbPBIHG。

Objectives：To improve wheat quality by gene genetic transformation. Methods：High molecular weight glutenin 12 subunit gene（HMW - GS 12） in wheat was amplified and cloned from wheat cultivar NE7742 genomic DNA by polymerase chain reaction.Results： Sequencing analysis showed the cloned fragment contained 1980 nueleotides and shared a homology of 99.5% and 99.7% respectively in DNA sequence and deduced amino acid sequence with the sequence published on genbank. By DNA processing, two plant expression vectors were constructed that one was endospermspecific expression vector and one constitutive expression vector pDNPPBIHG and pUbPBIHG, respectively.