【摘要】 以马铃薯栽培品种大西洋(Atlantic)试管苗叶片为材料,用Trizol试剂提取总RNA,通过RT-PCR方法获得马铃薯PPase基因的cDNA片段,将该片段反向连接到克隆载体pBluescriptSK+,酶切鉴定后进行基因测序,所得PPase基因片断为673bp,编码211个氨基酸,与已克隆的PPase基因同源性达到97.62%。将该PPase基因反向插入到表达载体pBI121、pBIC和pBIrd中,分别构建了组成型启动子CaMV35S、块茎特异表达启动子CIPP和低温诱导表达启动子rd29A驱动的反义PPase基因植物表达载体,以期利用转基因技术培育出块茎休眠期延长或缩短的马铃薯新种质。更多还原

【Abstract】 Total RNA was extracted from leaves of potato cultivar Atlantic using Trizol regent. Potato inorganic pyrophosphatase (PPase) gene cDNA was amplified using the reverse transcription polymerase chain reaction (RT-PCR) method and reversely cloned into pBluescript SK+ vector. The recombinant plasmid was identified by enzyme digestion. The sequence analysis showed that the PPase cDNA was 673 bp with 636 bp open reading frame (ORF) encoded a 211-amino acid polypeptide, which is 97.62% homology to the published sequence. The antisense PPase gene was inserted into expression vectors pBI121, pBIC and pBIrd to construct plant expression vectors, promoters of which are the constitutive promoter CaMV 35S, tuber-specific expression promoter CIPP and low temperature-inducible promoter rd29A, respectively, which would be applied to develop the potato germplasms with the long or short periodic characteristics of tuber dormancy.更多还原