论文部分内容阅读
分别用ＰｓｔⅠ、ＨｉｎｄⅢ和ＥｃｏＲⅠ三种限制性内切核酸酶切割提莫菲维小麦基因组ＤＮＡ，再与用相应酶处理的载体ｐＵＣ１１９连接，然后转化到大肠杆菌菌系ＤＨ５α之中。转化效率为１．２×１０４／μｇ。经α互补检测，初步筛选重组子。随机挑取单菌落分别培养，提取质粒，经琼脂糖凝胶电泳和斑点杂交，最后确定重组子，重组率为４３．８％。粗略地估计了克隆片段在基因组中的拷贝数
The genomic DNA of Timofeeve wheat was cut with three restriction endonucleases Pst I, Hind III and Eco RI, respectively, ligated with pUC119 vector treated with the corresponding enzymes, and then transformed into E. coli strain DH5α. The conversion efficiency was 1.2 × 10 4 / μg. After α complementary detection, preliminary screening of recombinant. Single colonies were randomly picked and cultured, the plasmid was extracted, agarose gel electrophoresis and dot blot hybridization, and finally to determine the recombinants, the recombination rate was 43.8%. A rough estimate of the copy number of the cloned fragment in the genome