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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
田　李^1,2, 张晓雯^1,2, 秦　松¹
1.中国科学院海洋研究所;2.中国科学院研究生院

摘要:

利用改良的SDS法提取钝顶螺旋藻基因组DNA ,经Sau3AI酶切,回收1～2 kb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(Escherichia coli)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。

关键词: 螺旋藻(Spirulina) 质粒文库 STK 克隆

DOI：

分类号:

基金项目:国家自然科学基金项目(30671065) ;中国科学院知识创新工程项目( KSCX2-YW-G-002 , KZCX2-YW-209)

Construction of DNA l ibrary of Spirulina and its appl ication in cloning genes

Abstract:

The genomic DNA isolated from Spirulina was digested by Sau3AI. The DNA fragments with length of 1～2 kb were cloned into plasmid vector pBR322 which was previously digested with BamHI and dephosphorylated with calf alkaline phosphatase. The recombinant plasmid was transformed into Escherichia coli Top10 competent cells and the genomic library was constructed. Using the anchor primer and degenerated primer,we obtain 5’end of STK in Spirulina from this genomic library.

Key words: Spirulina genomic library STK clone

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