Recombinational Cloning the Antibiotic Biosynthetic Gene Clusters in Linear Plasmid SCP1 of Streptomyces coelicolor A3(2)

Uso de un plásmido lineal para clonar genes de producción de antibióticos

Autores: 

1. Ran Zhang,
2. Haiyang Xia,
3. Qingyu Xu,
4. Fujun Dang,
5. Zhongjun Qin

The model organism Streptomyces coelicolor A3(2) harbors a 356-kb linear plasmid, SCP1. We report here development of a recombinational cloning method for deleting large segment from one telomere of SCP1 followed by replacing with the telomere of pSLA2 and sequentially inserting with the overlapping cosmids in vivo. The procedure depends on homologous recombination coupled with cleavage at telomere termini by telomere terminal protein. Using this procedure we cloned the 81-kb avermectin and the 76-kb spinosad biosynthetic gene clusters into SCP1. Heterologous expression of avermectin production in S. coelicolor was detected. These results demonstrate the utility of SCP1 for cloning large DNA segments such as antibiotic biosynthetic gene clusters.

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