Heterologous expression of Avermectins biosynthetic gene cluster by construction of a Bacterial Artificial Chromosome library of the producers.
Heterologous expression of Avermectins biosynthetic gene cluster by construction of a Bacterial Artificial Chromosome library of the producers.
复制标题
通过构建生产者细菌人工染色体文库异源表达阿维菌素生物合成基因簇。
DOI：
10.1016/j.synbio.2017.03.001
复制
发表时间：
2017-03
影响因子：
4.8
通讯作者：
Zhao C
中科院分区：
文献类型：
--
作者：
Deng Q;Zhou L;Luo M;Deng Z;Zhao C
阿维菌素是一类多酮类天然产物，在农业中被广泛用作驱虫剂。代谢工程和组合生物合成被广泛应用于提高阿维菌素的产量和创造新的阿维菌素衍生物，包括伊维菌素和多拉菌素。在体内对阿维菌素生产商阿维链霉菌进行遗传操作是劳动密集型和时间成本高的。阿维菌素生物合成基因簇的克隆和异源表达将使体外定制该基因簇成为可能。我们构建了白花链霉菌ATCC 31267细菌人工染色体文库，插入的DNA片段大小在100~130kb之间。从文库中筛选出5个携带阿维菌素生物合成基因簇AVE(81kb)的重组BAC克隆。然后，AVE在利维丹链霉菌中异源表达。从重组菌株的细胞提取液中检测到A2a、B1a和A1a三种阿维菌素成分。通过聚酮合成酶结构域的互换，为阿维菌素衍生物的开发提供了便利，为阿维菌素合成生物学研究提供了功能元件。
Avermectins, a group of polyketide natural products, are widely used as anthelmintics in agriculture. Metabolic engineering and combinatorial biosynthesis were extensively employed to improve Avermectins production and create novel Avermectin derivatives, including Ivermectin and Doramectin. It is labor intensive and time cost to genetically manipulate Avermectins producer Streptomyces avermitilis in vivo. Cloning and heterologous expression of Avermectins biosynthetic gene cluster will make it possible to tailor the cluster in vitro. We constructed a Bacterial Artificial Chromosome (BAC) library of S. avermitilis ATCC 31267 with inserted DNA fragments ranged from 100 to 130 Kb. Five recombinant BAC clones which carried the Avermectins biosynthetic gene cluster ave (81 Kb in size) were screened out from the library. Then, ave was hetero-expressed in S. lividans. Three Avermectin components, A2a, B1a and A1a were detected from the cell extracts of recombinant strains. It will facilitate the development of Avermectin derivatives by polyketide synthase domain swapping and provide functional element for Avermectins synthetic biology study.
登录
查看更多内容
影响因子：
3.9
作者：
Gonzalez-Ruiz A;Seaton RA;Hamed K
通讯作者：
Hamed K
鸟氨酸转氨甲酰酶 ArgK 在产生菜豆毒素的丁香假单胞菌的自卫中发挥双重作用。
影响因子：
4.6
作者：
Chen L;Li P;Deng Z;Zhao C
通讯作者：
Zhao C
dnd 基因对细菌中 DNA 进行硫代磷酸化
影响因子：
14.8
作者：
Wang, Lianrong;Chen, Shi;Dedon, Peter C.
通讯作者：
Dedon, Peter C.
影响因子：
2.7
作者：
Li, Meng;Chen, Zhi;Li, Jilun
通讯作者：
Li, Jilun
影响因子：
4.9
作者：
BURG, RW;MILLER, BM;OMURA, S
通讯作者：
OMURA, S