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Background  Mycobacteriophages are viruses that infect  Mycobacterium  hosts. A large collection of phages known to infect the same bacterial host strain –  Mycobacterium smegmatis  mc 2 155 – exhibit substantial diversity and characteristically mosaic architectures. The well-studied lytic mycobacteriophage D29 appears to be a deletion derivative of a putative temperate parent, although its parent has yet to be identified.    Results  Here we describe three newly-isolated temperate phages – Kerberos, Pomar16 and StarStuff – that are related to D29, and are predicted to be very close relatives of its putative temperate parent, revealing the repressor and additional genes that are lost in D29. Transcriptional profiles show the patterns of both lysogenic and lytic gene expression and identify highly-expressed, abundant, stable, small non-coding transcripts made from the P left  early lytic promoter, and which are toxic to  M. smegmatis .    Conclusions  Comparative genomics of phages D29, Kerberos, Pomar16 and StarStuff provide insights into bacteriophage evolution, and comparative transcriptomics identifies the pattern of lysogenic and lytic expression with unusual features including highly expressed, small, non-coding RNAs.    Electronic supplementary material  The online version of this article (10.1186/s12866-017-1131-2) contains supplementary material, which is available to authorized users.

Expression and evolutionary patterns of mycobacteriophage D29 and its temperate close relatives