Plasmid cloning vector pAS1

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

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435320, 935 29, 935 23, C12N 1500, C12N 100

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049257994

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 plasmic closely resembles the earlier constructed pKC30cII, also a regulatory protein which activates promoters for lysogenic development.

REFERENCES:
patent: 4578355 (1986-03-01), Rosenberg
Rosenberg, M. et al., in Methods in Enzymology, vol. 101, pp. 123-138, 1983.
Morrow, J. in Methods in Enzymology, vol. 68, pp. 3-24, 1979.
Shatzman and Rosenberg, "A Plasmid Cloning Vector for Inducible Overproduction of Proteins in Bacterial Cells," Miami Symposium, Jan. 14, 1982, Abstract, p. 98.
Shimatake and Rosenberg, Nature, vol. 292, No. 5819, pp. 128-132, Jul. 9, 1981.
Lewin, Gene Expression-3, John Wiley, 1977, pp. 352, 355, 371.
Kornberg, DNA Replication, W. H. Freeman and Co., 1980, pp. 539-540.
Backman et al., Cell, 13:65-71, Jan. 1978.
Old et al., Principles of Gene Manipulation, 2d ed., Univ. of California Press, 1981, p. 35+.

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