Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
 31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

To study genetic relationships between isolates of Ross River virus (RRV), an alphavirus with a chromosome of approximately 12,000 nucleotides, total high-molecular-weight RNA from RRV-infected baby hamster kidney (BHK) cells was transcribed into 32P-labeled, complementary DNA using reverse transcriptase and random calf-thymus DNA primers. The cDNA was digested with HaeIII or TaqI (restriction nucleases which cleave single-stranded DNA), and the restriction fragments separated on a standard DNA sequencing gel. The resulting HaeIII or TaqI restriction digest profiles mainly comprised virus-specific bands; cell RNAs were transcribed poorly. In reconstruction experiments, purified 49 S RRV genomic RNA and a 10-fold mass excess of mock-infected-cell RNA were reverse transcribed in the same reaction mix. Under these conditions there was no interference with the transcription of viral RNA sequences. When the level of viral RNA was lowered to one-hundredth that of cell RNA in the reaction mix, there was no qualitative change in restriction digest profiles. The procedure is rapid, simple, uses small amounts of 32P, does not require purification of virus or viral RNA, and permits cross-comparison between several virus strains on a single one-dimensional gel. The method should be applicable to other single-stranded RNA viruses of moderate genome complexity.
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PMID:Analysis of Ross River virus genomic RNA using HaeIII digests of single-stranded cDNA to infected-cell RNA and virion RNA. 241 9

Shoalwater Bay military training area (SWBTA), 2,713 km2 of land located 50-80 km north of Rockhampton, Queensland, Australia, is used by Australian and allied forces for training purposes. Between March 1998 and February 2000, monthly collections of mosquitoes at 15 sites were conducted using carbon dioxide-baited traps to study the seasonal occurrence of mosquitoes and Ross River virus (RRV) and Barmah Forest virus (BFV) in mosquitoes. A total of 72,616 mosquitoes, comprising 3,897 pools were collected, and 2,428 pools were tested using a reverse transcriptase-polymerase chain reaction. A total of 15 pools of mosquitoes were positive for virus, 10 RRV and five BFV. Blood meals from an additional 763 mosquitoes were tested by a gel diffusion assay, and the majority (96%) of those identified were from kangaroo, which was the most common mammal in the study area. The results indicate that Culex annulirostris Skuse and Ochlerotatus vigilax (Skuse) are the main vectors of RRV at SWBTA.
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PMID:Occurrence of Ross River virus and Barmah Forest virus in mosquitoes at Shoalwater Bay military training area, Queensland, Australia. 1498 54

To determine the presence of arboviruses in mosquito populations from major urban areas of eastern Australia, a total of 67,825 mosquitoes, representing -60 species, was collected and tested from Cairns, Brisbane, and Sydney between January 2005 and April 2008. Mosquito pools were screened by inoculation onto mosquito cell cultures and the detection of viral antigen using a panel of flavivirus and alphavirus monoclonal antibodies in an enzyme-linked immunosorbent assay. Suspect positive samples were confirmed using virus-specific real-time reverse transcriptase-polymerase chain reaction assays. No flaviviruses were detected, but 2 alphaviruses were isolated from mosquito pools collected from Cairns, with 1 Barmah Forest virus isolate from a pool of 100 Aedes vigilax and 1 Ross River virus isolate from a pool of 83 Verrallina carmenti. In addition, a single Aedes alternans collected from Sydney yielded an isolate most similar to Stretch Lagoon virus, a newly described virus from the genus Orbivirus. These results suggest that during the study, arboviruses were circulating at a low level in the areas sampled. The findings from this study will promote public health awareness of the risk of arboviruses in urban areas, leading to more informative public health campaigns to safeguard the Australian public.
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PMID:Arboviruses isolated from mosquitoes collected from urban and peri-urban areas of eastern Australia. 1985 16