摘要
以绛红色小单孢菌G1008基因组为模板,构建genY基因缺失的同源重组质粒pFY103,经接合转移导入绛红色小单孢菌G1008和GK1101(△genK),筛选获得genY基因缺失工程菌GY105(△genY)和工程菌GKY205(△genK+genY),发酵、提取并经质谱检测分析代谢产物.结果表明,工程菌GY105和GKY205主要积累西索米星、庆大霉素C1a和C2b.证明genY基因缺失阻断了庆大霉素X2到G418的转化,说明genY基因参与庆大霉素生物合成过程中绛红糖胺C-6’位甲基化.
The homologous recombination plasmid pFY103 with genY gene deletion was built by taking micromonospora purpura G1008 genome as the template.The micromonospora purpurea G1008 and GK1101（△genK）were guided into by conjugational transfer and genY gene deletion engineering bacteria GY105（△genY）and GKY205（△genK＋genY）were screened out.It fermented,extracted,and analyzed the metabolite through mass spectrometric detection.The result shows that engineering bacteria GY105 and GKY205 primarily accumulate sisomicin,Gentamycin C1 aand C2b,which proves that the gene deletion of genYgene blocks the transfer of gentamicin X2 into gentamicin G418,and explains the gene of genY is involved in the methylation of purpurosamine C-6＇in the biosynthesis process of gentamicin.
出处
《宁夏大学学报（自然科学版）》
CAS
2016年第1期85-89,共5页
Journal of Ningxia University(Natural Science Edition)
基金
国家自然科学基金资助项目(31070093)
国家"重大新药创制"专项基金资助项目(2012ZX09201101-008)
