Total DNA transcription in vitro: a procedure to detect highly repetitive and transcribable sequences with tRNA-like structures
Total DNAs from various animals were transcribed in vitro in a HeLa cell extract, and it was found that one to several discrete RNAs were transcribed by RNA polymerase III. With tortoise (Geoclemys reevessi) and newt (Cynops pyrrhogaster), distinct 6.5S and 8S RNAs were transcribed from these respective DNAs. Representative phage clones carrying the 6.5S and 8S RNA genes were isolated from genomic libraries of these animals, and the sequences of these genes were determined. The 5' parts of highly repetitive and transcribable sequences of tortoise and newt were found to have close resemblance to tRNA/sub 1//sup Lys/ (rabbit) gene (78% homology) and a tRNA/sup Glu/ (Drosophila) gene (74% homology, not counting the aminoacyl stem region), respectively. The homologies extended to secondary structures, homologous nucleotides being located on similar secondary structures. It is proposed that many, if not all, highly repetitive and transcribable sequences detected by total DNA transcription have specific tRNA genes as their progenitors.
- Research Organization:
- Univ. of Tsukuba, Ibaragi-ken, Japan
- OSTI ID:
- 6220466
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 83:2
- Country of Publication:
- United States
- Language:
- English
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HELA CELLS
DNA SEQUENCING
LABELLED COMPOUNDS
HYBRIDIZATION
AMPHIBIANS
BIOLOGICAL EVOLUTION
DNA
DROSOPHILA
ELECTROPHORESIS
GENES
IN VITRO
MOLECULAR BIOLOGY
PHOSPHORUS 32
RABBITS
TRANSCRIPTION
TRANSFER RNA
ANIMALS
AQUATIC ORGANISMS
ARTHROPODS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
DAYS LIVING RADIOISOTOPES
DIPTERA
FLIES
FRUIT FLIES
INSECTS
INVERTEBRATES
ISOTOPES
LIGHT NUCLEI
MAMMALS
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RNA
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques