Plasmids for production of human growth hormone or polypeptide a

Details Plasmids for production of human growth hormone or polypeptide a Plasmids for production of human growth hormone or polypeptide a

1994-11-14

1997-06-10

Martinell, James

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Bacteria or actinomycetales; media therefor

4353201, C12N 1500, C12N 1570

Patent

active

056374950

ABSTRACT:
An improved vector upon introduction into a suitable bacterial host containing the thermolabile repressor C.sub.I renders the host cell capable, upon increasing the temperature of the host cell to a temperature at which the repressor is destroyed, of effecting expression of a desired gene inserted into the vector and production of polypeptide encoded by the gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: a DNA sequence which contains the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site for binding antiterminator N protein produced by the host cell; a DNA sequence which contains a ribosomal binding site for rendering the mRNA of the desired gene capable of binding to ribosomes within the host cell; an ATG initiation codon or a DNA sequence which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a restriction enzyme site for inserting the desired gene into the vector in phase with the ATG initiation codon; and additionally a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell and a DNA sequence which contains a gene associated with a selectable or identifiable trait which is manifested when the vector is present in the host cell.

REFERENCES:
patent: 4578355 (1986-03-01), Rosenberg
patent: 4925799 (1990-05-01), Rosenberg
Rosenberg et al, Methods Enzymol. 101: 123 (1983).
R. Watson, et al., FEBS Letters, 150(1); 114-116 (1982).

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