Abstract:

In order to construct bacterial artificial chromosomes library of Streptomyces albulus var.wuyiensis,we extractd high molecular weight DNA(HMW-DNA) from Streptomyces albulus var.wuyiensis.Mycelium pellet of Streptomyces albulus var.wuyiensis was embeded in low melting point agarose.The agarose block was treated by lysozyme,proteinase K and restriction enzyme.Contour-clamped homogeneous electric field(CHEF) gel electrophoresis was performed for prepared DNA plugs.Electrophoresis analysis shows no degradation...

Key words: high molecular weight DNA, bacterial artificial chromosomes library, CHEF gel electrophoresis;

摘要：

为构建小白链霉菌武夷变种(Streptomyces albulus var. wuyiensis)的细菌人工染色体(bacterial artificial chromosomes,BAC)文库,提取了小白链霉菌武夷变种高分子量DNA(high molecular weight DNA,HMW-DNA)。用低熔点琼脂糖(low melting point agarose,LMP)包埋小白链霉菌武夷变种菌丝球,胶块经溶菌酶、蛋白酶K、两种限制性内切酶(BamHI、Hind III)处理后,使用箝位匀强电场(CHEF)凝胶电泳检测DNA片段大小。电泳结果显示提取HWM DNA完整,满足构建BAC文库的条件,确定了最适酶切条件为Hind III 0.2 U酶切10min,为小白链霉菌武夷变种功能基因的研究奠定了基础。

关键词: 高分子量DNA 细菌人工染色体 箝位匀强电场电泳