Nucleotide sequence of the 3'-nontranslatable region of mRNA of cow. cap alpha. /sub s/sub 1//-casein

Journal Article · Mon Jul 01 04:00:00 EDT 1985 · Dokl. Biol. Sci. (Engl. Transl.); (United States)

OSTI ID:5856447

Kyarshulite, D R; Zakhar'ev, V M; Gorodetskii, S I

In this paper, the authors make a determination of the nucleotide sequence of the 3'-nontranslatable region of cow ..cap alpha../sub s/sub 1//-casein mRNA and an analysis of the degree of homology with the corresponding region in rat mRNA. In subsequent analysis, the authors established that in the cloned sequence of cDNA (740 nucleotide pairs) there are no recognition sites for the restriction endonucleases MspI and PstI, and there is one site for the restriction endonuclease EcoRI. On the basis of restriction analysis and blot hybridization with /sup 32/P-labeled poly(T), the authors determined the orientation of cDNA in the DNA of the plasmid pK cas ..cap alpha../sub s/sub 1//-17. These data were used in selecting approaches to the determination of the nucleotide sequence. Labeled DNA fragments were obtained by hydrolysis of the plasmid DNA with the restriction endonuclease EcoRI, followed by introduction of the label at the ends with the aid of E. coli DNA polymerase I (large fragment) in the presence of (/sup 32/P)-dATP and three unlabeled dNTP. After supplementary hydrolysis of DNA with the restriction endonuclease PstI, the mixture was fractionated by electrophoresis in 5% polyacrylamide gel, and the labeled fragments corresponding to cDNA 9450 and 300 n.p. were extracted.

• Research Organization: N. I. Vavilov Institute of General Genetics, Moscow, USSR
• OSTI ID: 5856447
• Journal Information: Dokl. Biol. Sci. (Engl. Transl.); (United States), Journal Name: Dokl. Biol. Sci. (Engl. Transl.); (United States) Vol. 280:1-6; ISSN DKBSA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ATP
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CASEIN
CATTLE
CHEMICAL REACTIONS
CLONING
COWS
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
DNA
DNA-CLONING
DOMESTIC ANIMALS
ELECTROPHORESIS
ENZYMATIC HYDROLYSIS
ENZYMES
ESCHERICHIA COLI
ESTERASES
HYBRIDIZATION
HYDROLASES
HYDROLYSIS
ISOTOPES
LABELLED COMPOUNDS
LABELLING
LIGHT NUCLEI
LYSIS
MAMMALS
MESSENGER-RNA
MICROORGANISMS
NUCLEASES
NUCLEI
NUCLEIC ACIDS
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PHOSPHODIESTERASES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PROTEINS
RADIOENZYMATIC ASSAY
RADIOISOTOPES
RATS
RNA
RODENTS
RUMINANTS
SOLVOLYSIS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES