Comparative genomic analysis of mycobacteriophage Tweety: evolutionary insights and construction of compatible site-specific integration vectors for mycobacteria

• DOI: 10.1099/mic.0.2007/008904-0
• Publication Date: 2007-07-27T20:23:08Z
• Authors: Thuy T. Pham; Deborah Jacobs-Sera; Marisa L. Pedulla; Roger W. Hendrix; Graham F. Hatfull
• Keywords: Mycobacterium smegmatis; Lysogen

Mycobacteriophage Tweety is a newly isolated phage of Mycobacterium smegmatis. It has viral morphology with an isometric head and long flexible tail, forms turbid plaques from which stable lysogens can be isolated. The genome 58 692 bp in length, contains 109 protein-coding genes, shows significant but interrupted nucleotide sequence similarity the previously described mycobacteriophages Llij, PMC Che8. However, overall possesses mosaic architecture, gene products being related to other such as Che9d, Omega Corndog. A encoding integrase tyrosine-recombinase family located close centre genome, putative attP site been identified within short intergenic region immediately upstream int. This attP-int cassette was used construct new set integration-proficient plasmid vectors that efficiently transform both fast- slow-growing mycobacteria through integration at chromosomal locus containing tRNA(Lys) gene. These are maintained well absence selection completely compatible derived mycobacteriophage L5, enabling simple construction complex recombinants genes integrated simultaneously different positions.