PubMed · 4133183
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B J Poiesz, N Battula, L A Loeb. 1974-02-27. Zinc in reverse transcriptase.. https://doi.org/10.1016/s0006-291x(74)80282-2
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We developed a non-radioisotopic (non-RI) reverse transcriptase assay (RTA). The reverse transcriptase (RT) incorporates biotin-11-deoxyuridine-triphosphate (bio-dUTP) using a poly(rA) template hybridized with oligo(dT) primer that is immobilized on the surface of a 96-well microtiter plate. This assay is thus semi-automated by adapting it to an ELISA testing format. The incorporation of bio-dUTP was enhanced by adding cold dTTP to the reaction mixture, optimally in a molar ratio 4:1 (dTTP:bio-dUTP). This non-RI RTA is more sensitive than the conventional RI assay for the detection of purified Rous-associated virus 2 (RAV-2) and of human immunodeficiency virus type 1 (HIV-1) lysate. Because of its simple procedure, higher sensitivity and non-use of RI materials, the assay can be utilized not only for virological studies but also for routine safety screening of biological products for retroviral contamination.
erb-B gene is an oncogene from avian leukosis virus. The carcinogenic capability of erb-B is due to the close similarity of expressional proteins to the epidermal growth factor receptor (EGFr). In this study the amplification and the over-expression of erb-B were found in human oral squamous (cell carcinomas by DNA and RNA dot blot hybridization). Results showed that erb-B was correlated with the oral epidermoid tumors but not with group of non-epidermoid tumors in the head and neck region. Southern blotting analysis showed that there were rearrangement of erb-B gene in the tumor genosomes, which is perhaps one of the ways to activate the proto-oncogene of erb-B.
A study of the secondary structures of the 5'-leader RNA sequences of avian leukosis/sarcoma viruses was conducted using phylogenetic sequence alignment, theoretical structures calculated from base-pairing interactions involving the calculated minimal delta G values, and RNaseT1 sensitivity. The results suggest that all of the avian retroviral RNA leaders may be able to adopt similar conformations. Open reading frames in the leader RNAs may be positioned to facilitate viral activities such as translation and packaging of the genomic RNA into virus particles.