重组pET30a/hsBLyS质粒在BL21(DE3)菌株中的稳定性观察

Study on the heredity stability of a recombinant plasmid pET30a/hsBLyS in E.coli BL21(DE3)

将构建的含有pET30a/hsBLyS重组质粒的BL21 (DE3)菌种,在含硫酸卡那霉素的平板培养基划线连续传代至 100代 100代次菌种经显微观察,为典型的革兰氏阴性大肠杆菌,生化特性没有改变 提取0、50、100代质粒DNA限制性内切酶切割检查,酶切图谱没有改变,DNA测序未见hsBLyS基因变异 传代前后菌种诱导表达hsBLyS。

The strain of pET30a/hsBLyS/BL21 (DE3)had been subcultured consecutively from the original stock to the 100^(th) generation on LB plates with kanamycin.The digestion products of plasmid in the original ,the 50^(th) and the 100^(th) generation by NdeⅠand BamHⅠrestriction enzymes are same.As well,DNA sequences of the hsBLyS gene shows no differences among the generations.In addition to that,the original strain and the 100th generation strain have a similar hsBLyS expression level and these hsBLyS proteins show similar activites.