机构地区: 天津科技大学食品工程与生物技术学院
出 处: 《食品与发酵工业》 2004年第3期70-73,共4页
摘 要: 以耐高温α 淀粉酶生产菌地衣芽孢杆菌染色体DNA为模板 ,通过PCR扩增耐高温α 淀粉酶基因 ,将扩增产物 1 9kb的DNA片段插入到pUC1 9质粒中 ,再转化大肠杆菌JM1 0 9,通过在淀粉平板上形成透明圈等方法筛选到一株阳性克隆菌株JM1 0 9(pUAM) ,其表达产物可分泌到培养基中 ,除去菌体的发酵液中每 1 0 0mL酶活可达到 2 7个单位。将发酵上清液浓缩后用冷无水乙醇分级沉淀 ,所得样品进行SDS PAGE分析 ,得到分子质量为 60ku的目的蛋白带。 The heat-stable α-amylase gene was amplified from Bacillus licheniformis 020401 using PCR technique. The amplified 1.9*#kb DNA fragment was inserted into the vector pUC19 to yield the recombinant plasmid pUAM, which was cloned in E.coli JM109. The transter can secrete the α-amylase into the medium. The activity of the α-amylase reached 27 units/100*#mL of the cell-free supernatant of the culture. The cell-free supernatant of the culture was purified by cold alcohol. The molecular weight of the α-amylase was about 60 ku by SDS-PAGE.
关 键 词: 地衣芽孢杆菌 耐高温 淀粉酶基因 大肠杆菌 分泌机制 基因克隆
领 域: [生物学]