CAJ | 학술논문

以容纳DNA大片段的柯斯质粒Supercos-1为载体.构建了除虫链霉菌的基因组文库.对约1000个柯斯质粒上插入片段的两端进行测序,并利用全基因组序列的信息,构建了覆盖约91%基因组的有序排列的柯斯文库.利用入λ-Red高效DNA重组和大肠埃希菌-链霉菌接合转移技术,建立了除虫链霉菌的基因组遗传操作系统.运用该系统可以在除虫链霉菌工业菌株中进行高效、精确的基因敲除工作和连续的基因缺失研究,获得可以用来生产多拉菌素的工程菌株.
이용납DNA대편단적가사질립Supercos-1위재체.구건료제충련매균적기인조문고.대약1000개가사질립상삽입편단적량단진행측서,병이용전기인조서렬적신식,구건료복개약91%기인조적유서배렬적가사문고.이용입λ-Red고효DNA중조화대장애희균-련매균접합전이기술,건립료제충련매균적기인조유전조작계통.운용해계통가이재제충련매균공업균주중진행고효、정학적기인고제공작화련속적기인결실연구,획득가이용래생산다랍균소적공정균주.
A cosmid library of Streptomyces avermitilis genome was constructed by using vector Supercos-1. Sequencing of the insertion ends of nearly 1000 cosmids showed that they covered 91% genomic sequences of the strain. A genetic manipulation system was established by the λ-Red mediated recombination and conjugation from E. coli to Streptomyces. Genes of the strain were knocked out with high efficiency and site-preciseness, and even sequential gene knocking out was also practical. Several industrial strains with modifications of antibiotic biosynthetic pathways, producing doramectin for example, were obtained.