T4 DNA fragment as a stabilizer for proteins expressed by cloned

Details T4 DNA fragment as a stabilizer for proteins expressed by cloned T4 DNA fragment as a stabilizer for proteins expressed by cloned

1990-04-18

1991-11-12

Nucker, Christine

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Using a micro-organism to make a protein or polypeptide

4351721, 4351722, 4353201, 4352523, 43525233, 43525234, 536 27, 536 28, 536 29, 935 6, 935 33, 935 47, 935 48, 935 49, C12N 1500

Patent

active

050647571

ABSTRACT:
A DNA fragment isolated from the DNA of T-even bacteriophage which suppresses the degradation of abnormal proteins in bacterial cells is disclosed. The DNA fragment can be isolated by treating the DNA of T-even bacteriophage with a restriction enzyme to cleave the DNA into fragments, at least one of said fragments consisting essentially of a sequence of nucleotide bases which code for protein that functions to suppress degradation of abnormal proteins in bacterial cells, inserting the DNA fragments into cloning vehicles, such as plasmids, to produce hybrid cloning vehicles, transforming bacterial cells by introducing the hybrid cloning vehicles therein, and selecting the transformants which exhibit suppressed protein degradation mechanisms. The DNA fragment can then be isolated from the selected transformants. The stabilizing DNA fragment can be introduced into bacterial host cells to increase the yield of foreign proteins expressed by cloned DNA.

REFERENCES:
patent: 4237224 (1980-12-01), Cohen et al.
patent: 4336336 (1982-06-01), Silhavy et al.
Lee D. Simon & Kathleen Tomczak, "Bacteriophages Inhibit Degradation of Abnormal Proteins in E. coli", Nature, vol. 275, No. 5679, pp. 424-428, Oct. 5, 1978.
Lee D. Simon et al., "Hyperdegradation of Proteins in Escherichia coli rho Mutants", Proc. Natl. Acad. Sci. U.S.A., vol. 76, No. 4, pp. 1623-1627, Apr. 1979.
Humayun et al., The Single-Stranded DNA Phages (Denhardt, Ed.), Cold Spring Harbor, 1978, pp. 477-481.
Wais et al., Growth and Transformation of Phage T4 in Escherichia coli B/4, Salmonella, Aerobacter, Proteus, and Serratia, 1969, Virology 39, pp. 153-161.
Skorupski et al., A Bacteriophage T4 Gene Which Functions to Inhibit Escherichia coli Lon Protease, 1988, Journal of Bacteriology, pp. 3016-3024.
Richard J. Roberts, Recombinant DNA, Methods in Enzymology, vol. 68, 1979, pp. 27-35, 40.
Rupprecht et al., Conservation of capR (lon) DNA of Escherichia coli K-12 Between Distantly Related Species, 1983, Journal of Bacteriology, vol. 155, No. 2, pp. 910-914.
Benjamin Lewin, Plasmids and Phages, 1977, Gene Expression, p. 536.

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