Conjugation of φBT1-derived integrative plasmid pDZL802 in Amycolatopsis mediterranei U32

Bioengineered

Chen Li¹  Li Zhou¹  Xiaoming Ding¹  Ying Wang¹  Guoping Zhao² 

[1] State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, Chin;State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, Chin;Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Centre at Shanghai, Shanghai, Chin;CAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, Chin;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, Chin;

关键词: Amycolatopsis;    conjugal transfer system;    integrative plasmid;    metabolic engineering;    natural products;   

DOI  :  10.1080/21655979.2016.1270808

来源: Taylor & Francis

【 摘 要 】

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 × 10−5 CFU 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.

【 授权许可】

Unknown