Plasmids with conditional uncontrolled replication behavior

Details Plasmids with conditional uncontrolled replication behavior Plasmids with conditional uncontrolled replication behavior

1984-05-16

1989-02-21

Wiseman, Thomas G.

Chemistry: molecular biology and microbiology

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

Enzymatic production of a protein or polypeptide

435 70, 4351723, 4355171, 435320, 43525233, 935 29, 935 32, 935 92, 536 27, C12N 1500, C12N 120, C12P 2100, C12P 1934

Patent

active

048064718

ABSTRACT:
Plasmids useful as cloning vectors carrying an inserted regulatable promoter, transcription from the promoter regulating plasmid replication. Increased transcription leads to a substantially increased or uncontrolled plasmid copy number when host microorganisms containing the plasmid are cultivated under conditions securing such increased transcription. The regulatable promoter is preferably .lambda. P.sub.R, the activity of which is controlled by a temperature-sensitive .lambda. cl repressor. To obtain gene products of the plasmids, host microorganisms to which such plasmids have been transformed are desirably cultivated at about 30.degree. C. to secure a constant, low plasmid copy number during the seeding and multiplication stages of the microorganisms, after which the temperature is preferably shifted to about 36.degree.-42.degree. C. to obtain an uncontrolled plasmid copy number and an amplified amount of gene product.

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