λ Vectors for Stable Cloned Gene Expression

Biotechnology Progress - Tập 6 Số 4 - Trang 277-282 - 1990
Nandan Padukone1, S. Peretti1, David F. Ollis1
1Department of Chemical Engineering, North Carolina State University, Raleigh, North Carolina 27695‐7905

Tóm tắt

AbstractThe bacteriophage λ offers a unique opportunity concurrently to minimize segregational instability in recombinant systems by chromosomal integration of the cloned gene and to achieve high cloned gene expression during an abortive lytic phase. Lysis leads approximately to a 100‐fold amplification of the cloned gene. Cell lysis in the lytic state is blocked by a specific mutation (Sam), allowing the cell to maintain its integrity, and λ DNA packaging is blocked by other mutations (Wam, Eam) that keep cloned genes open to transcription. In the presence of these mutations, extremely high levels of cloned β‐galactosidase (more than 15% of total cell protein) have been obtained during abortive lysis from vectors found to be essentially 100% stable for over 75 generations in the lysogenic phase.

Từ khóa


Tài liệu tham khảo

10.1007/BF00393671

10.1007/BF00400172

Breton A. M., 1985, Transfer of plasmid RP4 to Myxococcus xanthus and evidence for its integration into the chromosome, J. Bacterial., 161, 523, 10.1128/jb.161.2.523-528.1985

10.1016/0168-1656(86)90009-X

10.1002/bit.260271014

10.1111/j.1749-6632.1986.tb26490.x

10.1038/nbt0384-251

Franklin N. C., 1971, The Bacteriophage Lambda, 621

10.1089/dna.1985.4.273

10.1038/nbt1288-1402

10.1007/BF00267194

Huynh T. V., 1984, DNA Cloning Techniques: A Practical Approach

10.1111/j.1749-6632.1981.tb14172.x

Maniatis T., 1982, Molecular Cloning: A Laboratory Manual, 440

McKenney K., 1981, Gene Amplification and Analysis, Vol. II: Structural Analysis of Nucleic Acids, 383

10.1016/0092-8674(80)90639-X

10.1002/j.1460-2075.1985.tb03989.x

10.1038/nbt0986-802

10.1007/BF00275963

10.1007/BF00267858

10.1038/nbt1086-901

10.1126/science.322281

10.1038/nbt0190-47

10.1016/0378-1119(81)90106-2

10.1016/0378-1119(83)90164-6

10.1002/bit.260320602

10.1002/bit.260270105

10.1016/0378-1119(83)90042-2

10.1016/0378-1119(87)90314-3

10.1002/bit.260330808

10.1007/BF00156608

10.1016/0378-1119(86)90367-7

Yamaguchi K., 1983, Copy Number Mutations (cop−) of the Plasmid Containing the Replication Origin (oriC) of the E. coli Chromosome: Lethal Effect of the cop Region onto a High‐Copy‐Number Vector on Host Cells, J. Bacteriol., 153, 550, 10.1128/jb.153.1.550-554.1983

10.1016/0378-1119(85)90120-9

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Biotechnology Progress

Tập 6 Số 4

277-282

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