title: Improved isolation of undersampled bacteriophages: finding of distant terminase genes.
abstract: Isolation and characterization of new environmental bacteriophages are performed for (1) analyzing microbial evolution and ecology and (2) delivering biological therapy. The sampling of environmental bacteriophages appears, however, to be limited by the procedure (usually liquid enrichment culture) used to propagate them. An alternative, less competitive procedure is developed here for the purpose of isolating new bacteriophages. This procedure involves extraction directly into and then propagation in a dilute agarose gel. Adaptations of this procedure are used to avoid repeated isolation of the same bacteriophage. Some newly isolated bacteriophages grow so poorly that they appear inaccessible to liquid enrichment culture. Four comparatively high titer bacteriophages were isolated and characterized by a genomic sequence survey. Some had genomes with extremely distant relationships to those of other bacteriophages, based on a tree built from the large terminase genes. These methods find novel genomes by rapidly isolating and screening diverse bacteriophages.
authors.affiliation: Department of Biochemistry, The University of Texas Health Science Center, San Antonio, TX 78229-3900, USA. serwer@uthscsa.edu
authors.forename: Philip
authors.initials: P
authors.lastname: Serwer
authors.forename: Shirley J
authors.initials: SJ
authors.lastname: Hayes
authors.forename: Shehnaz
authors.initials: S
authors.lastname: Zaman
authors.forename: Karen
authors.initials: K
authors.lastname: Lieman
authors.forename: Mandy
authors.initials: M
authors.lastname: Rolando
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Hardies
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics; D001435:Bacteriophages / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D003470:Culture Media; D004706:Endodeoxyribonucleases / Q000235:genetics*; D004783:Environmental Microbiology; D016679:Genome, Viral; D008969:Molecular Sequence Data; D010802:Phylogeny; D012685:Sepharose; D012689:Sequence Homology, Nucleic Acid
chemical_list: D003470:Culture Media; D012685:Sepharose; D004706:Endodeoxyribonucleases; C030894:terminase