caa a22 4500 605503508 CHVBK 20210128100611.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00253-014-6369-0 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6369-0 An efficient blue-white screening based gene inactivation system for Streptomyces [Elektronische Daten] [Pengwei Li, Jine Li, Zhengyan Guo, Wei Tang, Jianshan Han, Xiangxi Meng, Tingting Hao, Yaxin Zhu, Lixin Zhang, Yihua Chen] Streptomyces is studied intensively for its outstanding ability to produce bioactive secondary metabolites and for its complicated morphological differentiation process. A classical genetic manipulation system for Streptomyces has been developed and widely used in the community for a long time, using antibiotic resistance markers to select for double-crossover mutants. The screening process is always laborious and time-consuming. However, the lack of a suitable chromogenic reporter for Streptomyces has limited the use of color-based screening system to simplify the selection process for double-crossover mutants. In this study, a blue reporter system for Streptomyces has been established by mining an indigoidine synthetase gene (idgS) from Streptomyces lavendulae CGMCC 4.1386, leading to the development of a time-saving gene inactivation system for Streptomyces by simple blue-white screening. A series of Streptomyces suicide and temperature-sensitive plasmids containing the idgS reporter cassette were constructed and used successfully to inactivate genes in Streptomyces, allowing a simple and efficient screening method to differentiate the colonies for double-crossover (white) and single-crossover (blue) mutants. Inactivation of the putative γ-butyrolactone synthase gene afsA-y via the idgS-based blue-white screening method revealed that the paulomycin production is negatively controlled by afsA-y in Streptomyces sp. YN86. Springer-Verlag Berlin Heidelberg, 2015 Streptomyces nationallicence Indigoidine nationallicence Gene inactivation nationallicence Blue-white screening nationallicence Li Pengwei State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Li Jine State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Guo Zhengyan State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Tang Wei State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Han Jianshan State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Meng Xiangxi State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Hao Tingting State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Zhu Yaxin State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Zhang Lixin CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut Chen Yihua State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1923-1933 0175-7598 99:4<1923 2015 99 253 https://doi.org/10.1007/s00253-014-6369-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6369-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Li Pengwei State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 700 1- Li Jine State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 700 1- Guo Zhengyan State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 700 1- Tang Wei State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 700 1- Han Jianshan State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 700 1- Meng Xiangxi State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 700 1- Hao Tingting State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 700 1- Zhu Yaxin State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 700 1- Zhang Lixin CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut NATIONALLICENCE 700 1- Chen Yihua State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Chaoyang District, 100101, Beijing, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1923-1933 0175-7598 99:4<1923 2015 99 253