title: Conjugation of ϕBT1-derived integrative plasmid pDZL802 in Amycolatopsis mediterranei U32.
abstract: The genus Amycolatopsis is well known for its ability to produce antibiotics, and an increasing number of valuable biotechnological applications, such as bioremediation, biodegradation, bioconversion, and potentially biofuel, that use this genus have been developed. Amycolatopsis mediterranei is an industrial-scale producer of the important antibiotic rifamycin, which plays a vital role in antimycobacterial therapy. Genetic studies of Amycolatopsis species have progressed slowly due to the lack of efficient transformation methods and stable plasmid vectors. In A. mediterranei U32, electroporation and replicable plasmid vectors have been developed. Here, we establish a simple and efficient conjugal system by transferring integrative plasmid pDZL802 from ET12567 (pUZ8002) to A. mediterranei U32, with an efficiency of 4 × 10-5CFU per recipient cell. This integrative vector, based on the ϕBT1 int-attP locus, is a stable and versatile tool for A. mediterranei U32, and it may also be applicable to various other Amycolatopsis species for strain improvement, heterologous protein expression, and synthetic biology experiments.
authors.affiliation: a State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering , School of Life Sciences, Fudan University , Shanghai , China.
authors.forename: Chen
authors.identifier: http://orcid.org/0000-0001-8757-1153
authors.initials: C
authors.lastname: Li
authors.affiliation: a State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering , School of Life Sciences, Fudan University , Shanghai , China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhou
authors.affiliation: a State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering , School of Life Sciences, Fudan University , Shanghai , China.
authors.forename: Ying
authors.identifier: http://orcid.org/0000-0002-5841-3321
authors.initials: Y
authors.lastname: Wang
authors.affiliation: a State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering , School of Life Sciences, Fudan University , Shanghai , China.|b Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Centre at Shanghai , Shanghai , China.|c CAS Key Laboratory of Synthetic Biology , Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences , Shanghai , China.|d Department of Microbiology and Li Ka Shing Institute of Health Sciences , The Chinese University of Hong Kong, Prince of Wales Hospital , Shatin, New Territories, Hong Kong SAR , China.
authors.forename: Guoping
authors.initials: G
authors.lastname: Zhao
authors.affiliation: a State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering , School of Life Sciences, Fudan University , Shanghai , China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Ding
mesh_terms: D039903:Actinobacteria / Q000235:genetics* / Q000378:metabolism*; D003227:Conjugation, Genetic / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D012294:Rifamycins / Q000096:biosynthesis*; D014162:Transfection / Q000379:methods*; D014170:Transformation, Genetic / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D012294:Rifamycins
keywords: Amycolatopsis; conjugal transfer system; integrative plasmid; metabolic engineering; natural products