Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 2 de 2
Filter
Add more filters










Database
Language
Publication year range
1.
Biochim Biophys Acta ; 1814(9): 1107-12, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21586349

ABSTRACT

pV(VGJΦ), a single-stranded DNA binding protein of the vibriophage VGJΦ was subject to biochemical analysis. Here, we show that this protein has a general affinity for single-stranded DNA (ssDNA) as documented by Electrophoretic Mobility Shift Assay (EMSA). The apparent molecular weight of the monomer is about 12.7kDa as measured by HPLC-SEC. Moreover, isoelectrofocusing showed an isoelectric point for pV(VGJΦ) of 6.82 pH units. Size exclusion chromatography in 150mM NaCl, 50mM sodium phosphate buffer, pH 7.0 revealed a major protein species of 27.0kDa, suggesting homodimeric protein architecture. Furthermore, pV(VGJΦ) binds ssDNA at extreme temperatures and the complex was stable after extended incubation times. Upon frozen storage at -20°C for a year the protein retained its integrity, biological activity and oligomericity. On the other hand, bioinformatics analysis predicted that pV(VGJΦ) protein has a disordered C-terminal, which might be involved in its functional activity. All the aforementioned features make pV(VGJΦ) interesting for biotechnological applications.


Subject(s)
Bacteriophages/chemistry , DNA-Binding Proteins/chemistry , Vibrio cholerae/chemistry , Viral Proteins/chemistry , Computational Biology , Isoelectric Point , Molecular Weight , Protein Multimerization
2.
J Bacteriol ; 191(18): 5873-6, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19617366

ABSTRACT

The native product of open reading frame 112 (orf112) and a recombinant variant of the RstB protein, encoded by Vibrio cholerae pathogen-specific bacteriophages VGJphi and CTXphi, respectively, were purified to more than 90% homogeneity. Orf112 protein was shown to specifically bind single-stranded genomic DNA of VGJphi; however, RstB protein unexpectedly bound double-stranded DNA in addition to the single-stranded genomic DNA. The DNA binding properties of these proteins may explain their requirement for the rolling circle replication of the respective phages and RstB's requirement for single-stranded-DNA chromosomal integration of CTXphi phage dependent on XerCD recombinases.


Subject(s)
Bacteriophages/metabolism , DNA-Binding Proteins/metabolism , Inovirus/metabolism , Vibrio cholerae/virology , Viral Proteins/metabolism , Virus Integration , Attachment Sites, Microbiological , Bacteriophages/classification , Bacteriophages/genetics , DNA/metabolism , DNA, Single-Stranded/metabolism , DNA-Binding Proteins/genetics , Electrophoresis, Polyacrylamide Gel , Gene Expression Regulation, Viral , Inovirus/classification , Inovirus/genetics , Open Reading Frames/genetics , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Viral Proteins/genetics
SELECTION OF CITATIONS
SEARCH DETAIL
...