title: Characterization of the RstB2 protein, the DNA-binding protein of CTXϕ phage from Vibrio cholerae.
abstract: The low abundant protein RstB2, encoded in the RS2 region of CTXϕ, is essential for prophage formation. However, the only biochemical activity so far described is the single/double-stranded DNA-binding capacity of that protein. In this paper, a recombinant RstB2 (rRstB2) protein was overexpressed in E. coli with a yield of 58.4 mg l(-1) in shaken cultures, LB broth. The protein, purified to homogeneity, showed an identity with rRstB2 by peptide mass fingerprinting. The apparent molecular weight of the RstB2 native protein suggests that occurs mostly as a monomer in solution. The monomers were able of reacting immediately upon exposure to DNA molecules. After a year of storage at -20 °C, the protein remains biologically active. Bioinformatics analysis of the amino acid sequence of RstB2 predicts the C-end of this protein to be disordered and highly flexible, like in many other single-stranded DNA-binding proteins. When compared with the gVp of M13, conserved amino acids are found at structurally or functionally important relative positions. These results pave the way for additional studies of structure and molecular function of RstB2 for the biology of CTXϕ.
authors.affiliation: National Center for Scientific Research, Ave 25 and 158, Cubanacán, Playa, PO Box 6214, Havana, Cuba, alina.falero@cnic.edu.cu.
authors.forename: Alina
authors.initials: A
authors.lastname: Falero
authors.forename: Karen
authors.initials: K
authors.lastname: Marrero
authors.forename: Sonia
authors.initials: S
authors.lastname: Trigueros
authors.forename: Rafael
authors.initials: R
authors.lastname: Fando
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016415:Sequence Alignment; D014734:Vibrio cholerae / Q000821:virology*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D014764:Viral Proteins