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

Pham, TT and Jacobs-Sera, D and Pedulla, ML and Hendrix, RW and Hatfull, GF (2007) Comparative genomic analysis of mycobacteriophage Tweety: Evolutionary insights and construction of compatible site-specific integration vectors for mycobacteria. Microbiology, 153 (8). 2711 - 2723. ISSN 1350-0872

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Abstract

Mycobacteriophage Tweety is a newly isolated phage of Mycobacterium smegmatis. It has a viral morphology with an isometric head and a long flexible tail, and forms turbid plaques from which stable lysogens can be isolated. The Tweety genome is 58 692 bp in length, contains 109 protein-coding genes, and shows significant but interrupted nucleotide sequence similarity with the previously described mycobacteriophages Llij, PMC and Che8. However, overall the genome possesses mosaic architecture, with gene products being related to other mycobacteriophages such as Che9d, Omega and Corndog. A gene encoding an integrase of the tyrosine-recombinase family is located close to the centre of the genome, and a putative attP site has been identified within a short intergenic region immediately upstream of int. This Tweety attP-int cassette was used to construct a new set of integration-proficient plasmid vectors that efficiently transform both fast- and slow-growing mycobacteria through plasmid integration at a chromosomal locus containing a tRNAL Lys gene. These vectors are maintained well in the absence of selection and are completely compatible with integration vectors derived from mycobacteriophage L5, enabling the simple construction of complex recombinants with genes integrated simultaneously at different chromosomal positions. © 2007 SGM.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Pham, TT
Jacobs-Sera, Ddjs@pitt.eduDJS0000-0001-7838-3257
Pedulla, ML
Hendrix, RWrhx@pitt.eduRHX
Hatfull, GFgfh@pitt.eduGFH
Date: 1 August 2007
Date Type: Publication
Journal or Publication Title: Microbiology
Volume: 153
Number: 8
Page Range: 2711 - 2723
DOI or Unique Handle: 10.1099/mic.0.2007/008904-0
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 1350-0872
MeSH Headings: Amino Acid Sequence; Attachment Sites, Microbiological--genetics; DNA, Viral--chemistry; DNA, Viral--genetics; Genetic Vectors; Integrases--genetics; Lysogeny; Microscopy, Electron, Transmission; Molecular Sequence Data; Mycobacteriophages--genetics; Mycobacteriophages--physiology; Mycobacteriophages--ultrastructure; Mycobacterium smegmatis--virology; Open Reading Frames; Plasmids--genetics; Recombination, Genetic; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; Transformation, Bacterial; Virion--ultrastructure
Other ID: NLM PMC2884959
PubMed Central ID: PMC2884959
PubMed ID: 17660435
Date Deposited: 05 Dec 2012 16:37
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/16638

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