Comparison of efficiency of use of early and late promoters of T7 phage for β-galactosidase genie expression in Escherichia coli
DOI:
https://doi.org/10.7124/bc.000250Abstract
The model system for comparison of the efficiency of use of early and late promoters of T7 phage for the β-galactosidase gene expression has been constructed. The following strategy of construction has been chosen. The gene of T7 phage RNA-polymerase has been inserted into DNA of phage cloning vector. The model gene z of lac operon of E. coli (EcoRI-Sall digest of pLZ56) has been inserted into DNA of plasmid GEM-1 (firm «Promega») which contains phage T7 late promoter. The cloned pLZ56 fragment of DNA contains also S-D sites and A3 promoter specifically recognized by E. coli RNA-polymerase. Some features of b-galactosidase synthesis during transcription of the z gene under the control of the late promoter of phage T7 and under the control of A3 promoter are shown.References
Tabor S, Richardson CC. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985;82(4):1074-8.
Studier FW, Moffatt BA. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986;189(1):113-30.
Moffatt BA, Dunn JJ, Studier FW. Nucleotide sequence of the gene for bacteriophage T7 RNA polymerase. J Mol Biol. 1984;173(2):265-9.
Charnay P, Louise A, Fritsch A, Perrin D, Tiollais P. Bacteriophage lambda-E. coli K12 vector-host system for gene cloning and expression under lactose promoter control. II. DNA fragment insertion at the vicinity of the lac UV5 promoter. Mol Gen Genet. 1979;170(2):171-8.
Davanloo P, Rosenberg AH, Dunn JJ, Studier FW. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1984;81(7):2035-9.
Fuerst TR, Niles EG, Studier FW, Moss B. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1986;83(21):8122-6.
Miller D. Determination of lactose operon enzymes. Moscow, Mir, 1976; 436 p.
Chernykh SI, Glushakova LG, Kordium VA. Phage-dependent beta-galactosidase synthesis by Escherichia coli cells. Mikrobiol Zh. 1979;41(5):479-83.
Maniatis T, Fritsch EF, Sambrook J. Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Lab. press, 1982 545 p.
Moir A, Brammar WJ. The use of specialised transducing phages in the amplification of enzyme production. Mol Gen Genet. 1976;149(1):87-99.
Glushakova LG, Chernykh SI, Stel'mashenko LN, Kordium VA. Beta-galactosidase super synthesis induced by phage Xplac5cl857some amber-mutants. Molekulyarnaya biologiya (Kiev). 1982; Is. 30:37-41.