Plasmid library for the transcription of RNA probes complementary to the entire genome of the human immunodeficiency virus type 1 (HIV-1). NLM AIDSLINE Important note: Information in this article was accurate in 1990. The state of the art may have changed since the publication date.

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Plasmid library for the transcription of RNA probes complementary to the entire genome of the human immunodeficiency virus type 1 (HIV-1).

Biochem Cell Biol. 1989 Sep;67(9):510-5. Unique Identifier : AIDSLINE MED/90027690
Roy C; Knaak C; Garvie P; Limmer B; Campione-Piccardo J; Molecular Virology Laboratory, Health and Welfare Canada, Ottawa,; Ont.

Abstract: A plasmid (pBH10R3) containing a 9-kb Sst I fragment of HIV-1 (clone BH-10) inserted in pSP64, an in vitro expression vector, has been used for the transcription of anti-sense HIV-1 RNA. With this system, the transcripts obtained in vitro were not usually full length (1 to 2 kb long) and they predominantly span the 3' end ORF and ENV regions of the viral genome. We have rearranged the HIV-1 genomic sequences with respect to the SP6 promoter in the pSP64 vector and have obtained a series of new constructs allowing the expression in vitro of RNA transcripts complementary to other regions in the HIV-1 genome, including the 5' end of the ENV region as well as the TAT, POL, and GAG regions. In fact, the combined use of these constructs as templates for in vitro transcription allows the production of RNA probes spanning the entire viral genome. Compared with the 1- to 2-kb probes mentioned above, the combined use of such probes results in a several-fold increase in the sensitivity of molecular hybridization for the detection of HIV-1 nucleic acid sequences. Also, these constructs enable the preparation of RNA probes that have the potential to detect restriction polymorphisms throughout the HIV-1 genome.
Keywords: *Gene Library *Genes, Viral Genetic Vectors HIV-1/*GENETICS Nucleic Acid Hybridization Plasmids/*GENETICS *RNA Probes Support, Non-U.S. Gov't Templates *Transcription, Genetic JOURNAL ARTICLEKWDgenelibraryKWDgenes,viralgeneticvectorshiv-1/KWDgeneticsnucleicacidhybridizationplasmids/KWDgeneticsKWDrnaprobessupport,non-uKWDsKWDgov'ttemplatesKWDtranscription,geneticjournalarticle
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Copyright © 1990 - National Library of Medicine. Reproduced under license with the National Library of Medicine, Bethesda, MD.

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