Biopolym. Cell. 2001; 17(2):160-165.
Molecular and Cell Biotechnologies
Isolation of clones sensitive to bacteriophage λ from the phage resistant Escherichia coli strain
- Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
A method of selection of the phage λ sensitive clones from the population of the bacteriophage λ resistant cells using the phage λcI857ApRTcR has been theoretically formulated and experimentally approved. The isolation was realized as follows. The phage λ resistant E. coli strain was infected by phage λ, which introduces antibiotic-resistance markers into the cell. The sensitive to phage λ vir and lysing at 43 В°C clones were isolated on the selective medium. The frequency of occurrence of such cells in the population was 10–6 . The presence of ts-mutation in the phage cI gene has allowed to cure the cells from phage λcI857ApRTcR during cultivation of these cells under non-selective conditions at increased temperature and to obtain the phage A sensitive clones originated from resistant E. coli strain. The method can also be used for the estimation of the E. coli populations heterogeneity by a given feature.
 Moir A, Brammar WJ. The use of specialised transducing phages in the amplification of enzyme production. Mol Gen Genet. 1976;149(1):87-99.
 Drew RE, Clarke PH, Brammar WJ. The construction in vitro of derivatives of bacteriophage lambda carrying the amidase genes of Pseudomonas aeruginosa. Mol Gen Genet. 1980;177(2):311-20.
 AS. USSR N 1593232 Al. A method of producing ?-galactosidase. VA Kordyum, SI Chernykh, IYu Slavchenko, VG Korobko. 15.05.1990.
 Maniatis T, Fritsch EF, Sambrook J. Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Lab, 1982; 545 p.
 Slavchenko IIu, Chernykh SI, Kordium VA. [The use of the mutant phage lambda cI857 ApRTcRN- for isolation of cells lacking the prophage lambda]. Mol Gen Mikrobiol Virusol. 1985;(11):42-4.
 Pat. USA 4,784,952. Conferred susceptibility to lambda phage in non-coliform prokaryotic hosts. Robert A. Ludwig, Gert E. de Vries. Publ. 15.11.88.
 Chernykh SI, Stel'mashenko LN, Kordium VA. [Characteristics of the expression of the Ap- and Tc-genes of plasmid origin in lambda vectors]. Genetika. 1984;20(3):382-8.
 Kravchenko VV, Guileva IP, Shamin VV, Kulichkov VA, Dobrynin VN, Filippov SA, Chuvpilo SA, Korobko VG. Construction of a tandem of artificial genes encoding human leukocyte interferon and its expression as a part of polycistronic template with coupled translation. Bioorg Khim. 1987; 13 (9):1186-93.
 Kashyap SK, Bystrov NS, Boldyreva EF, Polyakova IA, Severtsova IV, Korobko VG. Chemical-enzymatic synthesis and cloning in Escherichia coli of genes coding for human granulocyte colony stimulating. Bioorg Khim. 1992; 18(1):71-7