Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (12): 168-172.doi: 10.13560/j.cnki.biotech.bull.1985.2014.12.028

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Identification and Molecular Characterization of a Phage from Streptomyces fradiae

Bai Yu, Xu Chunyan, Su Jianyu   

  1. (Key Laboratory of Ministry of Education for Conservation and Utilization of Special Biological Resources in the Western, School of Life Science, Ningxia University, Yinchuan 750021)
  • Received:2014-04-25 Online:2014-12-08 Published:2014-12-12

Abstract: It was to identify and explore the molecular characterization of a phage SF1, which was isolated from Streptomyces fradiae fermentation broth during the failed industrial production of tylosin. Phage SF1 was characterized by observing the shape of plaque, titering on double-layer agar culture plate. The morphology of SF1 was observed by the electron microscopic observation after negative staining. The sensitivity of the phage to chloroform and isopropyl alcohol was tested to detect its envelope. The structural proteins analysis of phage SF1 was carried out by SDS-PAGE. Restriction fragment length polymorphism(RFLP)analysis of genomic DNA was further performed to characterize the phage. Results showed that the plaque(11-15 mm in diameter)is neat and transparent. SF1 has a polyhedral head(about 24-30 nm in diameter)and a long tail(about 52-64 nm long and 3-5 nm wide). It is insensitive to chloroform and isopropyl alcohol. The genome of SF1 consists of double-stranded DNA and is approximately 35 kb. Capsid protein analysis shows a higher level of 14 major protein bands ranging from 14.8-59.5 kD, which is presumed to be the structural proteins of SF1.

Key words: Streptomyces fradiae, Phage, Tylosin