Genetic reagents for generating plasmids containing multiple cop

Details Genetic reagents for generating plasmids containing multiple cop Genetic reagents for generating plasmids containing multiple cop

1980-12-02

1983-09-06

Wiseman, Thomas G.

Chemistry: molecular biology and microbiology

Miscellaneous

435 68, 435 70, 435 91, C12N 100, C12P 2100, C12P 2104, C12P 1934

Patent

active

044030363

ABSTRACT:
Genetic reagents for generating plasmids containing multiples of a DNA segment are prepared by modification of the restriction sites of plasmid rings. The modified plasmids permit cloning of DNA segments which can be recovered with sticky ends that are complementary but not rotationally equivalent. Such segments will polymerize in a head-to-tail conformation. The plasmid rings with modified restriction sites are also used in linear form to obtain head-to-tail joining of multiple DNA segments into plasmid rings for further cloning and/or expression of the DNA segment-directed protein. The critical restriction site sequences of the modified plasmid rings can be prepared as a reagent which permits the sequence to be introduced into any plasmid. The reagents have utility in preparing multiples of protein-forming genes, and in preparing large amounts of homogeneous DNAs which can themselves be used as reagents.

REFERENCES:
patent: 4293652 (1981-10-01), Cohen
Sadler et al., Gene, 3, 211-232 (1978).
Sadler et al., Gene, 8, 279-300 (1980).
Lewin; Gene Expression-3, John Wiley & Sons, New York, 1977, p. 291.
Roberts; Methods in Enzymol. 68, 27 (1979).
Hohn; Methods in Enzymol. 68, 299 (1979).
Bolivar et al.: Gene 2, 95 (1977).

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