+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Insertion of bacteriophage lambda into the deo operon of Escherichia coli K-12 and isolation of plaque-forming lambdadeo+ transducing bacteriophages



Insertion of bacteriophage lambda into the deo operon of Escherichia coli K-12 and isolation of plaque-forming lambdadeo+ transducing bacteriophages



Journal of Bacteriology 136(2): 668-681



A procedure has been devised to isolate plaque-forming lambda cI857S7 transducing bacteriophage which carry the internal promoter, P3, of the deo operon of Escherichia coli and the deoB and deoD genes, while lacking the deoP and cytP promoters of the same operon, in order to study, specifically, regulation at the P3 site. This has been accomplished by selecting for the insertion of bacteriophage lambda into the deoA gene in a strain deleted for the normal lambda attachment site (delta att lambda) and isolating from this lysogen lambda spi- and lambda EDTAr phage. Among these, lambda pdeoB+D+ phage were identified by their transducing abilities. From in vivo enzyme induction experiments performed on a delta deo strain lysogenized with such phage, they were shown to carry the P3 promoter while lacking the deoP and cytP promoters. A lambdapdeo B+D+ phage phage was used to lysogenize a deo+ delta att lambda strain, integration of lambda occurring within the region of homology, and, from a heat-induced lysate of this strain, a plaque-forming lambda+ phage carrying the complete deo operon was obtained. Phage lambda was also inserted into the deoB and deoD genes and into the tdk gene. By isolating lambdaspi- and lambdaEDTAr phage from the deo::(lambda) mutants and determining which bacterial genes they carried and whether they retained the int gene of lambda, it was found that lambda had inserted into deoD with the same orientation as lambda inserted into attlambda, whereas lambda inserted into deoA and deoB had the opposite orientation. Deletions extending from the site of lambda insertion into the bacterial chromosome were isolated by selecting for heat-resistant revertants. These confirmed the order of markers to be deo-serB-trpR-thr and also placed a locus, msp, determining sensitivity or resistance of male strains to male-specific phages, between trpR and thr. For some reason unknown, but which may be related to the orientation of the lambda prophages, short deletions rendering the bacterium Ser- Thr+ were of much lower frequency from the deoD::(lambda) lysogen than from the other two lysogens. From an examination of the residual deoD enzyme levels in deoB::(lambda) mutants, it was deduced that there may be two promoter sites within the deoB::(lambda) mutants, it was deduced that there may be two promoter sites within the deoB gene, transcription from one of these being sufficient to account for the noncoordinate nature of the induction of deoB and deoD gene products.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 040456889

Download citation: RISBibTeXText

PMID: 361716


Related references

Isolation of specialized transducing bacteriophage lambda carrying genes of the L-arabinose operon of Escherichia coli B/r. Journal of Bacteriology 123(3): 1043-1054, 1975

Isolation and characterization of lambda b221poriCasnA, a plaque-forming specialized transducing phage carrying the origin of replication of the Escherichia coli chromosome. Molecular and General Genetics 178(2): 381-389, 1980

The isolation and characterization of plaque-forming arabinose transducing bacteriophage lambda. Journal of Molecular Biology 95(3): 395-407, 1975

Characterization of a defective lambda bacteriophage transducing the threonine operon of Escherichia coli K12. Molecular and General Genetics 132(1): 41-48, 1974

Isolation of specialized lambda transducing bacteriophages for flagellar genes (fla) of Escherichia coli K-12. Journal of Virology 22(3): 654-661, 1977

Isolation and characterization of nondefective transducing lambda bacteriophages carrying fla genes of Escherichia coli K-12. Journal of Bacteriology 130(2): 736-745, 1977

Defective and plaque-forming lambda transducing bacteriophage carrying penicillin-binding protein-cell shape genes: genetic and physical mapping and identification of gene products from the lip-dacA-rodA-pbpA-leuS region of the Escherichia coli chromosome. Journal of Bacteriology 143(2): 569-581, 1980

Isolation and characterization of lambda transducing bacteriophages for the su1+ (supD minus) amber suppressor of Escherichia coli. Journal of Bacteriology 122(1): 120-128, 1975

Isolation of specialized transducing bacteriophages carrying deletions of the regulatory region of the Escherichia coli K-12 tryptophan operon. Journal of Bacteriology 128(1): 283-289, 1976

Directed transposition of the arabinose operon: a technique for the isolation of specialized transducing bacteriophages for any Escherichia coli gene. Journal of Molecular Biology 44(1): 117-127, 1969

Physical characterization of the escherichia coli dna c region carried by a plaque forming lambda dna c transducing phage. Experientia 39(2): 187-188, 1983

Isolation of a lambda transducing bacteriophage carrying the relA gene of Escherichia coli. Journal of Bacteriology 127(2): 917-922, 1976

Isolation of a specialized transducing bacteriophage lambda carrying the polC locus of Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 71(7): 2614-2617, 1974

Isolation and characterization of a lambda transducing bacteriophage carrying the cysE gene of Escherichia coli K-12. Australian Journal of Biological Sciences 33(5): 605-612, 1980

Transposition of the lac region to the gal region of the Escherichia coli chromosome: isolation of lambda-lac transducing bacteriophages. Journal of Bacteriology 108(1): 5-9, 1971