【摘要】 我们将从地衣芽孢杆菌(Bacillus licheni-formis)克隆的约1.68kb的耐高温α-淀粉酶基因构建成表达载体,并转入根癌农杆菌。以马铃薯栽培品种“杂交荷兰7号”块茎圆盘为外植体,按本实验室建立起的再生实验系统及杨美珠等的方法进行转化。采取共培养、芽的诱导、芽的选择再生三步方法获得抗性芽。将抗性芽通过先诱导生根壮苗,再进行卡那霉素筛选,最后再诱导生根的方法得到可能的转基因植株。对部分可能的转基因植株按改进的王广立等的PCR简单快速鉴定转基因植物的方法进行检测,株号102001、102607、110402均可见到特异性片段的存在。参照张振清及王福荣等的方法对这些植株进行耐高温α-淀粉酶活力测定,这些植株具有相对较强的耐高温α-淀粉酶活性。实验结果表明,耐高温α-淀粉酶基因可能已转入上述植物基因组中,并获表达。更多还原

【Abstract】 The thermostableα-amylase gene cloned from Bacillus lichemiformis was reconstructed into an expression vector pAMY721M under the CaMV35S promoter. The vector was transferred into A. tumerfaciens ABI. The thermostable a-amylase gene was transferred into Solanum tuberosum L. via Agrobacterium mediation according to the revised method of ZHAO Shu Juan et al (1997) and YANG Mei Zhu et al (1992). Shoots were induced and regenerated on MSmedium with 2mg/L ZT, 0. 1mg/L IAA and 100mg/L kanamycin. Putative transformants were selected with kanamycin and roots induced on MS medium with 0. 15mg/L IAA. PCR analysis and thermostable a-amylase activity assay were done to identify the transgenic plantlets. A-mong them, 102001, 102607, and 110402 were showed to have a higher a-amylase activity than untransformed control.更多还原