scholarly journals A map of the cleavage sites for endonuclease AvaI in the chromosome of bacteriophage lambda

1977 ◽  
Vol 163 (3) ◽  
pp. 503-509 ◽  
Author(s):  
S G Hughes
Keyword(s):  
Linear Order ◽  
Bacteriophage Lambda ◽  
Cleavage Sites ◽  
Endonuclease Ecori ◽  
Fragmentation Patterns ◽  
Lambda Dna

The linear order of nine fragments generated by the action of endonuclease AvaI on the DNA of bacteriophage lambda was determined from the altered fragmentation patterns of bacteriophages containing known deletions and of hybrids of bacteriophages lambda and phi80. Digestion of 5′-terminally 32P-labelled bacteriophage-lambda DNA was used to identify the terminal fragments. Measurement of relative fragment lengths permitted rough mapping of the endonuclease-AvaI cleavage sites relative to the ends of the bacteriophage-lambda chromosome. The fragment order was confirmed and the map refined by analysis of the fragmentation of derivative phages containing single cleavage sites for endonuclease EcoRI.

scholarly journals The nucleotide sequences recognized by endonucleases AvaI and AvaII from Anabaena variabilis

1980 ◽  
Vol 185 (1) ◽  
pp. 65-75 ◽  
Author(s):  
S G Hughes ◽  
K Murray
Keyword(s):  
Nearest Neighbor ◽  
Bacteriophage Lambda ◽  
Nucleotide Sequences ◽  
Anabaena Variabilis ◽  
Cleavage Sites ◽  
Independent Evidence ◽  
Nearest Neighbor Analysis ◽  
The Individual ◽  
Lambda Dna

Determination of the 5'-terminal sequences flanking all the individual cleavage sites for endonuclease AvaI in bacteriophage-lambda DNA has shown that this enzyme recognizes the hexanucleotide sequences: (Formula: see text), This sequence is cut as shown by the arrows to give single-stranded 5'-tetranucleotide protrusions (cohesive ends). Endonucleases SmaI, XhoI and XmaI recognize different symmetrical subsets of this sequence and provide independent evidence for the occurrence of these subsets at particular endonuclease-AvaI cleavage sites in the bacteriophage-lambda genome. Further evidence for this structure came from the demonstration that DNA fragments generated by endonuclease AvaI can be ligated to form a discrete set of larger molecules and from nearest-neighbor analysis which showed that cytosine residues occurred at the 3'-side of cleavage points. The observation that endonuclease AvaII recognized a subset of the sites recognized by AsuI [Hughes, Bruce & Murray (1979) Biochem. J. 185, 59-63[led to the deduction that AvaII recognize the pentanucleotide sequence: (Formula: see text), and breaks internucleotide bonds at the positions indicated by the arrows.

scholarly journals Inhibition of transcription starting from bacteriophage lambda pR promoter during the stringent response in Escherichia coli: implications for lambda DNA replication.

1995 ◽  
Vol 42 (2) ◽  
pp. 233-239 ◽  
Author(s):  
A Szalewska-Pałasz ◽  
G Wegrzyn
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Dna Replication ◽  
Transcriptional Activation ◽  
Plasmid Dna ◽  
Bacteriophage Lambda ◽  
Stringent Response ◽  
Lacz Gene ◽  
Relative Level ◽  
Lambda Dna

Replication of lambda plasmid DNA is halted in amino acid-starved wild type (stringent) strains whereas it proceeds in relA (relaxed) mutants. The only transcription which could be important in lambda plasmid DNA replication in amino acid-starved Escherichia coli cells is that starting from the pR promoter. Using a fusion which consists of the lacZ gene under the control of bacteriophage lambda pR promoter we found that transcription starting from this promoter was inhibited during the stringent, but not the relaxed, response in E. coli. We confirmed our conclusion by estimating the relative level of the pR transcript by RNA-DNA hybridization. We propose that decreased transcription from the pR promoter which serves as transcriptional activation of ori lambda is responsible for inhibition of lambda plasmid replication during the stringent response. The results presented in this paper, combined with our recent findings (published elsewhere), indicate that the transcriptional activation of ori lambda may be a main regulatory process controlling lambda DNA replication not only during the relaxed response but also in normal growth conditions.

SphI restriction map of bacteriophage lambda DNA

Gene ◽  
1981 ◽  
Vol 16 (1-3) ◽  
pp. 321-323 ◽  
Author(s):  
Christoph Kessler ◽  
Georg Nesch ◽  
Ruth Brack
Keyword(s):  
Bacteriophage Lambda ◽  
Restriction Map ◽  
Lambda Dna
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