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λ Vectors for Stable Cloned Gene Expression
Author(s) -
Padukone N.,
Peretti S. W.,
Ollis D. F.
Publication year - 1990
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp00004a008
Subject(s) - lytic cycle , lysogenic cycle , biology , gene , bacteriophage , microbiology and biotechnology , recombinant dna , lysis , dna , genetics , escherichia coli , virus
The 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.