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海洋科学  2008 

钝顶螺旋藻基因组质粒文库的构建及其在获得功能基因上的应用

, PP. 55-60

Keywords: 螺旋藻(Spirulina),质粒文库,STK,克隆

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Abstract:

利用改良的SDS法提取钝顶螺旋藻基因组DNA,经Sau3AI酶切,回收1~2kb大小的片段,与BamHI(Sau3AI的同尾酶)酶切并去磷酸化后的pBR322质粒载体相连接,转入大肠杆菌(Escherichiacoli)Top10细胞,构建了螺旋藻基因组质粒文库。根据pBR322载体上多克隆位点两侧序列设计锚定引物,根据丝状蓝藻丝氨酸/苏氨酸激酶(STK)保守序列设计简并引物,以螺旋藻质粒文库为模板,“巢式”扩增出了STK基因部分序列。螺旋藻基因组质粒文库的构建为功能基因研究提供了极大方便。

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