Phagmids and genetic engineering: analysis of cloned gene libraries

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

OSTI ID:5841818

Mel'nikov, A A; Fodor, I

Phagmids are bi-replicon DNA molecules which, depending on the conditions, can form phage particles and lyse E. coli cells or be maintained in the cell in the plasmid state on account of a plasmid replicator. The authors suggest a new method for the selection of genes from cloned gene libraries, created on the basis of lambda phage. Phages from individual transparent plaques were reproduced, the DNA isolated, and the structure of the DNA was analyzed using restriction endonucleases and the method of hybridization. The authors used a fragment of the interferon A gene with which recombination was performed, as well as DNA fragments used for hybridization. The main evidence that clones containing interferon genes were selected by this method consists of the fact that recombination in vivo was performed with the 3'-end of the DNA of the interferon A gene, while DNA-DNA hybridization in the clones revealed the 5'-terminal sequences of the DNA of the gene. Hybridization of the EcoRI-BglII fragment of (/sup 32/P)-DNA of interferon A, both isolated from polyacrylamide gel and cloned in the vector M13mp8, showed that in five (lambda I2, I7, I8, I9, I11), of the ten selected phages, there are 5'-terminal fragments of the DNA of the interferon gene. The clones lambda I7, lambda I9, and lambda I11, have the same structure according to the data of restriction and hybridization analyses. The clones lambda I4, lambda I5, and lambda I6, are also identical and hybridize only with the 3'-terminal sequence of interferon A DNA.

• Research Organization: Institute of the Biochemistry and Physiology of Microorganisms, Pushchino, USSR
• OSTI ID: 5841818
• 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

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59 BASIC BIOLOGICAL SCIENCES
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CELL CONSTITUENTS
CHEMICAL REACTIONS
CLASSIFICATION
CLONING
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
DNA
DNA SEQUENCING
DNA-CLONING
ENZYMATIC HYDROLYSIS
ENZYMES
ESCHERICHIA COLI
ESTERASES
GENES
GENETIC ENGINEERING
GROWTH FACTORS
HYBRIDIZATION
HYDROLASES
HYDROLYSIS
INTERFERON
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LABELLING
LIGHT NUCLEI
LYMPHOKINES
LYSIS
MICROORGANISMS
MITOGENS
MOLECULAR STRUCTURE
NUCLEASES
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PLASMIDS
PROTEINS
RADIOENZYMATIC ASSAY
RADIOISOTOPES
SOLVOLYSIS
STRUCTURAL CHEMICAL ANALYSIS
TRACER TECHNIQUES