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Closed Access item Overproduction, purification, and characterization of DNA-binding protein P19 of bacteriophage PRD1
|Authors:||Pakula, Tiina M.|
Gutiérrez Armenta, Crisanto
Olkkonen, Vesa M.
Bamford, Dennis H.
|Keywords:||Protection of single-stranded DNA against nuclease P1, Phage λ pL promoter, Protein purification, Gel-shift assay|
|Citation:||Gene 126(1): 99-104 (1993)|
|Abstract:||The early protein, P 19, of bacteriophage PRD1 was purified after overexpression of the cloned gene, XIX, in Escherichia coli DH5α cells. The purified protein binds as multimers to single-stranded DNA (ssDNA), and with a lower affinity to double-stranded DNA (dsDNA), without sequence-specificity. Two distinct P19-ssDNA complexes were discovered in gel- mobility-shift assays at different protein:DNA ratios. P 19 was capable of fully protecting ssDNA against nuclease P1. Electron microscopy of protein P19-ssDNA complexes showed DNA molecules which were extensively coated with protein and whose contour length was clearly reduced by P 19 binding. The results suggest that P 19 binds to ssDNA with moderate cooperativity and are consistent with the DNA being wrapped around the P 19 multimers.|
|Publisher version (URL):||http://dx.doi.org/10.1016/0378-1119(93)90595-T|
|Appears in Collections:||(CBM) Artículos|
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