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Analysis of transcripts from predicted open reading frames of the Musca domestica salivary gland hypertrophy virus

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¹ E-mail: nzs2000{at}gmail.com

The Musca domestica salivary gland hypertrophy virus (MdSGHV) is a large dsDNA virus that infects and sterilizes adult houseflies. The transcriptome of this newly described virus was accomplished by rapid amplification of cDNA 3'-ends (3'-RACE) and reverse transcriptase polymerase chain reactions (RT-PCR). Direct sequencing of 3'-RACE products revealed 78 poly (A) transcripts containing 95 of the 108 putative ORFs. An additional six ORFs not amplified by 3'-RACE were detected by RT-PCR. Only seven of the 108 putative ORFs were not amplified by either 3'-RACE or RT-PCR. A series of 5'-RACE reactions were conducted on selected ORFs that were identified by 3'-RACE to be transcribed in tandem (tandem transcripts). In the majority of cases, only the downstream ORFs were detected as single transcripts and as components of the tandem transcripts. The only exception was the upstream ORF MdSGHV084, which was differentially transcribed as a single transcript at one and two days post-infection (dpi) and as a tandem transcript (MdSGHV084/085) at 2 dpi. Transcriptome analysis of MdSGHV detected splicing in the 3' untranslated region (3'-UTR) and extensive heterogeneity in the polyadenylation signals and cleavage sites. In addition, 21 overlapping sense-antisense transcripts were found. Finally, sequencing the 3'-RACE products without cloning served as an alternative approach to detect both 3'-UTRs and transcript variants of this large DNA virus.

Received 12 December 2008; accepted 17 January 2009.