Plasmid cloning vector pAS1

Details Plasmid cloning vector pAS1 Plasmid cloning vector pAS1

1983-01-12

1986-03-25

Wiseman, Thomas G.

Chemistry: molecular biology and microbiology

Miscellaneous

4351723, 435 68, 935 11, 935 29, C12N 100, C12N 1500, C12P 2100

Patent

active

045783551

ABSTRACT:
A plasmid cloning vector containing both transcriptional and translational regulatory sequences derived from the bacteriophage lambda genome was constructed to achieve high level expression of prokaryotic and eukaryotic genes. The system utilizes a plasmid vehicle carrying the strong, regulatable lambda promoter, P.sub.L, and host lysogens into which this vector can be stabily transformed. The lysogen synthesizes sufficient repressor (cI) to control P.sub.L expression and thereby stabilize plasmids which carry such a highly efficient promoter. Use of a temperature sensitive repressor permits simple, rapid induction of P.sub.L transcripts at any given time. Efficient transcription of essentially any coding sequence is assured by providing the phage lambda antitermination factor, N, and a site on the transcription unit for its utilization (Nut site). This pAS1 plasmid closely resembles the earlier constructed pKC30cII, also a regulatory protein which activates promoters for lysogenic development.

REFERENCES:
Bernard et al., Gene, 5:59 (1979).
Kornberg, DNA Replication, W. H. Freeman & Comp. pp. 539-540, 1980.
Old et al., Principles of Gene Manipulation, 2d. Ed., University of California Press, pp. 35-38, 1981.
Backman, et al., Cell, vol. 13, pp. 65-71, 1978.
Shatzman et al., "A Plasmid Cloning Vector for Inducible Overproduction of Proteins in Bacterial Cells", Miami Symposium, Abstract, p. 98, Jan. 14, 1982.
Shimatake et al., Nature, vol. 292, pp. 128-132, 1981.
Su et al., J. Biol. Chem, vol. 257, vol. 15, pp. 9128-9134, 1982.
Lewin, Gene Expression--3, John Wiley, pp. 352-355 and 371, 1977.

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