Plasmid vector with temperature-controlled gene expression

Journal Article · Sat Feb 01 00:00:00 EST 1986 · Sov. J. Bioorg. Chem. (Engl. Transl.); (United States)

OSTI ID:5347804

Kravchenko, V V; Yamshchikov, V F; Pletnev, A G

In plasmid pBR327, a fragment 169 b.p. long including promotor p/sub 3/ of the bla gene has been deleted. The deletional derivative so obtained (pSP2) has been used to construct a recombinant plasmid bearing a fragment of phage lambda DNA with the p/sub R/ promotor and the gene of the temperature-sensitive repressor cI. It has been shown that the plasmid vector so constructed (pCE119) with promotor cR performs repressor-cI-controlled transcription of the bla gene, as a result of which induction for an hour at 42/sup 0/C leads to an almost 100-fold increase in the amount of product of the bla gene as compared with that at 32/sup 0/C. The possibility of the use of plasmid cPE119 for the expression of other genes has been demonstrated for the case of the semisynthetic ..beta..-galactosidase gene of E. coli. In this case, on induction of the cells with recombinant plasmid pCEZ12 for 3 hours at 42/sup 0/C, a 300-fold increase in the amount of active ..beta..-galactosidase, as compared with that at 32/sup 0/C, was observed. It is important to point out that under these conditions (at 42/sup 0/C), at least 99% of the cells containing the plasmid retain the phenotype lacZ/sup +/, which indicates the stability of the proposed vector system

• Research Organization: All-Union Scientific-Research Institute of Molecular Biology, Novosibirsk (USSR)
• OSTI ID: 5347804
• Journal Information: Sov. J. Bioorg. Chem. (Engl. Transl.); (United States), Vol. 11:4; Other Information: Translated from Bioorganicheskaya Khim; 11: No. 4, 523-533(Apr 1985)
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PLASMIDS
BIOSYNTHESIS
GENE RECOMBINATION
RADIOENZYMATIC ASSAY
STRUCTURE-ACTIVITY RELATIONSHIPS
BACTERIOPHAGES
DNA POLYMERASES
ELECTROPHORESIS
ENZYME ACTIVITY
ESCHERICHIA COLI
GALACTOSIDASE
GENES
LABELLED COMPOUNDS
LABELLING
NUCLEOTIDES
PHENOTYPE
PHOSPHATES
PHOSPHORUS 32
RECOMBINANT DNA
THERMOREGULATION
TRANSCRIPTION
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CELL CONSTITUENTS
CONTROL
DAYS LIVING RADIOISOTOPES
DNA
ENZYMES
GLYCOSYL HYDROLASES
HYDROLASES
ISOTOPES
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
O-GLYCOSYL HYDROLASES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PARASITES
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
RADIOISOTOPES
SYNTHESIS
TEMPERATURE CONTROL
TRANSFERASES
VIRUSES
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