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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Bacterias que degradan el iboprufeno

 El ibuprofeno es un antiinflamatorio no esteroideo (AINE), utilizado frecuentemente como antipirético y para el alivio sintomático del dolor de cabeza (cefalea), dolor dental (odontalgia), dolor muscular o mialgia, molestias de la menstruación (dismenorrea), dolor neurológico de carácter leve y dolor postquirúrgico. También se usa para tratar cuadros inflamatorios, como los que se presentan en artritis, artritis reumatoide (AR) y artritis gotosa. En el último nº de Microbiology (SGM) se ha publicado un trabajo en el que se describe un microorganismo que degrada el iboprufeno.

Abstract

Sphingomonas Ibu-2 has the unusual ability to cleave the acid side chain from the pharmaceutical ibuprofen and related arylacetic acid derivatives to yield corresponding catechols under aerobic conditions via a previously uncharacterized mechanism. Screening a chromosomal library of Ibu-2 DNA in Escherichia coli EPI300 allowed us to identify one fosmid clone (pFOS3G7) that conferred the ability to metabolize ibuprofen to isobutylcatechol.

Iboprufeno

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