scholarly journals Cycle Sequencing of Filamentous Phage DNA Using a Biotinylated Primer and ΔTaq DNA Polymerase

BioTechniques ◽  
1997 ◽  
Vol 23 (1) ◽  
pp. 125-127 ◽  
Author(s):  
Jianping Qiu ◽  
Huanying Zhou ◽  
Joseph F. Aceto ◽  
Thomas Kieber-Emmons
Keyword(s):  
Dna Polymerase ◽  
Filamentous Phage ◽  
Cycle Sequencing ◽  
Phage Dna

scholarly journals Deep-sea vent phage DNA polymerase specifically initiates DNA synthesis in the absence of primers

2017 ◽  
Vol 114 (12) ◽  
pp. E2310-E2318 ◽  
Author(s):  
Bin Zhu ◽  
Longfei Wang ◽  
Hitoshi Mitsunobu ◽  
Xueling Lu ◽  
Alfredo J. Hernandez ◽  
...  
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deep Sea ◽  
Phage Dna

scholarly journals UV- and MMS-induced mutagenesis of lambdaO(am)8 phage under nonpermissive conditions for phage DNA replication.

2003 ◽  
Vol 50 (4) ◽  
pp. 921-939 ◽  
Author(s):  
Joanna Krwawicz ◽  
Anna Czajkowska ◽  
Magdalena Felczak ◽  
Irena Pietrzykowska
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Excision Repair ◽  
Protein S ◽  
Dna Lesions ◽  
E Coli ◽  
Phage Dna ◽  
Induced Mutagenesis ◽  
C Proteins ◽  
Inducible Protein

Mutagenesis in Escherichia coli, a subject of many years of study is considered to be related to DNA replication. DNA lesions nonrepaired by the error-free nucleotide excision repair (NER), base excision repair (BER) and recombination repair (RR), stop replication at the fork. Reinitiation needs translesion synthesis (TLS) by DNA polymerase V (UmuC), which in the presence of accessory proteins, UmuD', RecA and ssDNA-binding protein (SSB), has an ability to bypass the lesion with high mutagenicity. This enables reinitiation and extension of DNA replication by DNA polymerase III (Pol III). We studied UV- and MMS-induced mutagenesis of lambdaO(am)8 phage in E. coli 594 sup+ host, unable to replicate the phage DNA, as a possible model for mutagenesis induced in nondividing cells (e.g. somatic cells). We show that in E. coli 594 sup+ cells UV- and MMS-induced mutagenesis of lambdaO(am)8 phage may occur. This mutagenic process requires both the UmuD' and C proteins, albeit a high level of UmuD' and low level of UmuC seem to be necessary and sufficient. We compared UV-induced mutagenesis of lambdaO(am)8 in nonpermissive (594 sup+) and permissive (C600 supE) conditions for phage DNA replication. It appeared that while the mutagenesis of lambdaO(am)8 in 594 sup+ requires the UmuD' and C proteins, which can not be replaced by other SOS-inducible protein(s), in C600 supE their functions may be replaced by other inducible protein(s), possibly DNA polymerase IV (DinB). Mutations induced under nonpermissive conditions for phage DNA replication are resistant to mismatch repair (MMR), while among those induced under permissive conditions, only about 40% are resistant.

scholarly journals The TolQRA Proteins Are Required for Membrane Insertion of the Major Capsid Protein of the Filamentous Phage f1 during Infection

1998 ◽  
Vol 180 (7) ◽  
pp. 1723-1728 ◽  
Author(s):  
Eva Marie Click ◽  
Robert E. Webster
Keyword(s):  
Capsid Protein ◽  
Phage Particle ◽  
Cytoplasmic Membrane ◽  
Major Capsid Protein ◽  
Infection Process ◽  
Dna Transfer ◽  
Filamentous Phage ◽  
Membrane Insertion ◽  
Phage Dna ◽  
Phage Capsid

ABSTRACT Infection of Escherichia coli by the filamentous bacteriophage f1 is initiated by interaction of the end of the phage particle containing the gene III protein with the tip of the F conjugative pilus. This is followed by the translocation of the phage DNA into the cytoplasm and the insertion of the major phage capsid protein, pVIII, into the cytoplasmic membrane. DNA transfer requires the chromosomally encoded TolA, TolQ, and TolR cytoplasmic membrane proteins. By using radiolabeled phages, it can be shown that no pVIII is inserted into the cytoplasmic membrane when the bacteria contain null mutations in tolQ, -R and -A. The rate of infection can be varied by using bacteria expressing various mutant TolA proteins. Analysis of the infection process in these strains demonstrates a direct correlation between the rate of infection and the incorporation of infecting bacteriophage pVIII into the cytoplasmic membrane.

Terminal Cross-linking of DNA Catalyzed by an Enzyme System Containing DNA Ligase, DNA Polymerase, and Exonuclease of Bacteriophage T7

1974 ◽  
Vol 52 (6) ◽  
pp. 525-535 ◽  
Author(s):  
Paul Sadowski ◽  
Allison McGeer ◽  
Andrew Becker
Keyword(s):  
Dna Polymerase ◽  
Enzyme System ◽  
Maximal Activity ◽  
Cross Linking ◽  
Dna Ligase ◽  
Escherichia Coli Cells ◽  
Phage Dna ◽  
Phage T7 ◽  
Cross Links ◽  
T7 Exonuclease

An enzymatic activity that terminally cross-links T7 DNA has been detected in extracts from phage T7-infected Escherichia coli cells. This activity required the functional products of gene 1.3 (T7 DNA ligase), gene 5 (T7 DNA polymerase), and gene 6 (T7 exonuclease) as well as Mg2+ ions, the four deoxytriphosphates, and ATP. The cross-linking system could be reconstituted with purified ligase, polymerase, and exonuclease, and maximal activity required the presence of all three enzymes. The system was active on phage DNA which possessed unique termini but not on DNA with non-unique ends.

scholarly journals Appropriate Expression of Filamentous Phage f1 DNA Replication Genes II and X Requires RNase E-Dependent Processing and Separate mRNAs

1998 ◽  
Vol 180 (12) ◽  
pp. 3245-3249 ◽  
Author(s):  
Robert J. Kokoska ◽  
Deborah A. Steege
Keyword(s):  
Dna Replication ◽  
Mrna Stability ◽  
Continuous Production ◽  
Filamentous Phage ◽  
Overlapping Genes ◽  
Rnase E ◽  
Gene Encoding ◽  
E Coli ◽  
Phage Dna ◽  
Encoding Genes

ABSTRACT The products of in-frame overlapping genes II and X carried by the filamentous phage f1 genome are proteins with required but opposing functions in phage DNA replication. Their normal relative levels are important for continuous production of phage DNA without killing infected Escherichia coli hosts. Here we identify several factors responsible for determining the relative levels of pII and pX and that, if perturbed, alter the normal distribution of phage DNA species in infected hosts. Translation of the two proteins is essentially relegated to separate mRNAs. The mRNAs encoding genes II and X are also differentially sensitive to cleavage dependent onrne, the gene encoding the only E. coliendo-RNase known to have a global role in mRNA stability. Whereas pII levels are limited at the level of mRNA stability, normal pX levels require transcription in sufficient amounts from the promoter for the smaller mRNA encoding only pX.

Sign in / Sign up

Export Citation Format

Share Document