Expression vectors containing lambdapl promoter, T.sub.1 T.sub.2

Details Expression vectors containing lambdapl promoter, T.sub.1 T.sub.2 Expression vectors containing lambdapl promoter, T.sub.1 T.sub.2

1992-01-14

1992-09-29

Schwartz, Richard A.

Chemistry: molecular biology and microbiology

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

Bacteria or actinomycetales; media therefor

4353201, 935 45, 935 60, 935 29, C12N 1570, C12N 1573, C12N 121

Patent

active

051513641

ABSTRACT:
An improved vector upon introduction into a suitable host containing the thermolabile repressor C.sub.I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site; a first restriction enzyme site permitting replacement of the ribosomal binding site which follows thereafter; a ribosomal binding site for transcribing mRNA; and ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzyme site for inserting the gene in phase with the ATG codon; a T.sub.1 T.sub.2 rRNA transcription termination sequence; a DNA sequence which contains an origin of replication capable of autonomous production in the host of at least 400 constitutive copies of the vector; and either a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host cell or the fragment cI.sup.434 on which is included the gene for the repressor protein and its associated promoter and operator. The distance between the 3' end of P.sub.L O.sub.L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs.
Plasmids have been constructed from the vectors and used to produce bovine growth hormones.

REFERENCES:
patent: 4578355 (1986-03-01), Rosenberg

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