高级检索
李辉, 温淑平, 洪文荣. aprK基因敲除的氨甲酰妥布霉素工程菌的构建[J]. 中国药科大学学报, 2013, 44(4): 368-373. DOI: 10.11665/j.issn.1000-5048.20130416
引用本文: 李辉, 温淑平, 洪文荣. aprK基因敲除的氨甲酰妥布霉素工程菌的构建[J]. 中国药科大学学报, 2013, 44(4): 368-373. DOI: 10.11665/j.issn.1000-5048.20130416
shu
LI Hui, WEN Shuping, HONG Wenrong. Construction of carbamoyltobramcin-producing engineering S.tenebrarius by disruption of aprK gene[J]. Journal of China Pharmaceutical University, 2013, 44(4): 368-373. DOI: 10.11665/j.issn.1000-5048.20130416
Citation: LI Hui, WEN Shuping, HONG Wenrong. Construction of carbamoyltobramcin-producing engineering S.tenebrarius by disruption of aprK gene[J]. Journal of China Pharmaceutical University, 2013, 44(4): 368-373. DOI: 10.11665/j.issn.1000-5048.20130416
shu

aprK基因敲除的氨甲酰妥布霉素工程菌的构建

Construction of carbamoyltobramcin-producing engineering S.tenebrarius by disruption of aprK gene

    摘要
  • 摘要: 以温敏型穿梭质粒pKC1139为基础，克隆安普霉素辛二糖合成酶基因aprK上下游序列作为同源交换臂，构建用于敲除aprK的重组质粒pBK5。pBK5转化E.coli ET12567后经接合转移导入黑暗链霉菌Tt-49，得到单交换菌株ST315。ST315经松弛传代后通过影印培养与PCR筛选得到aprK阻断突变株ST316。ST316发酵效价约1 500 ug/mL，发酵产物经TLC分析，发现安普霉素的生物合成被阻断，得到了一株主要产氨甲酰妥布霉素的工程菌。

     

    Abstract: To construct a genetic engineering S. tenebrarius for production of carbamoyltobramycin. Recombinant plasmid pBK5 derivated from pKC1139 was constructed for disrupting gene aprK. pBK5 was introduced into S. tenebrariusTt-49 by conjugation. The single crossover mutant ST315 was obtained and cultivated for several rounds of sporulation in the absence of erythromycin. A desired double crossover mutant ST316 was achieved by PCR. TLC analysis indicated that ST316 produced high-yield carbamoyltobramycin, in which apramycin biosynthesis was blocked and the yield of carbamoyltobramycin was about 1 500 ug/mL.

     

    • HTML全文
    • 参考文献(0)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回

访问量：