Promotion of markerless deletion of the actinorhodin biosynthetic gene cluster in Streptomyces coelicolor.
The standard gene disruption and replacement to delete the actinorhodin biosynthetic gene cluster (Act) in Streptomyces coelicolor was inefficient, and the polymerase chain reaction-targeting of the cosmid could efficiently delete the Act, but still was a time-consuming procedure for markerless gene replacement. By using optimal Streptomyces codons, we synthesized a sceS gene encoding identical amino acid sequence as the chromosome rare-cutting meganuclease I-sce I of the Saccharomyces cerevisiae mitochondria. Expression of sceS gene in S. coelicolor resulted in promotion of homologous recombination and subsequently, successful achieved markerless deletion of the Act. The sceS system may be useful for the sequential markerless deletions of chromosomal segments in Streptomyces.
Lu
Zhiqun
Z
Shanghai Institute of Plant Physiology and Ecology, The Chinese Academy of Sciences, China.
Xie
Pengfei
P
Qin
Zhongjun
Z
D000880:Anthraquinones / Q000378:metabolism; D003360:Cosmids / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D005798:Genes, Bacterial / Q000235:genetics*; D005813:Genes, Synthetic / Q000235:genetics; D008957:Models, Genetic; D005810:Multigene Family / Q000235:genetics*; D009154:Mutation; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D014170:Transformation, Genetic; D000095622:Benzoisochromanequinones
D000880:Anthraquinones; D015252:Deoxyribonucleases, Type II Site-Specific; D029701:Saccharomyces cerevisiae Proteins; C066166:AI4 protein, S cerevisiae; C013390:actinorhodin; D000095622:Benzoisochromanequinones