Isolation and characterization of Micromonospora phage PhiHAU8 and development into a phasmid.
PhiHAU8, a temperate Micromonospora phage, which is capable of infecting Micromonospora sp. strains 40027 and A-M-01, was isolated. The PhiHAU8 virion has a polyhedral head and a flexible tail and has a small genome (ca. 42.5 kb) with double-stranded DNA and cohesive ends. PhiHAU8 was most stable at 4 degrees C in Difco nutrient broth within a pH range of 6 to 12. PhiHAU8 plaque formation on Micromonospora sp. strain 40027 was optimal with 32 mM Ca(2+) and 30 mM Mg(2+). A lysogen, LXH8, was isolated from turbid plaques, and a phasmid derivative that functions as a lambda cosmid vector in Escherichia coli and as a phage in Micromonospora sp. strain 40027 was constructed. Pulsed-field gel electrophoresis of AseI-digested total DNA showed that PhiHAU8 DNA integrates into the 500-kb AseI fragment of Micromonospora sp. strain 40027.
Li
Xiaohua
X
Bio-X Life Science Research Center, Shanghai Jiaotong University, Shanghai 200030, People's Republic of China.
Zhou
Xiufen
X
Deng
Zixin
Z
D001435:Bacteriophages / Q000235:genetics / Q000302:isolation & purification*; D003360:Cosmids; D004279:DNA, Viral / Q000235:genetics; D008242:Lysogeny; D008847:Micromonospora / Q000821:virology*
D004279:DNA, Viral