Molecular cloning, expression, and characterization of dnaK in Streptococcus pneumoniae.
DnaK is known to be highly conserved in all species and is a major immunogen in Streptococcus pneumoniae. To elucidate the role of dnaK in S. pneumoniae, dnaK was cloned in Escherichia coli using a homologous dnaK probe generated by PCR. The His-tagged DnaK was overexpressed in soluble form and purified from E. coli. Alignment of the deduced DnaK amino acid sequence from nucleotide sequences of the cloned dnaK revealed high homology with DnaK analogs in E. coli (53%) and Staphylococcus aureus (73%). However, anti-pneumococcal DnaK antiserum did not crossreact with DnaK analogs in E. coli, S. aureus and human cells suggesting that pneumococcal DnaK might be a good candidate as a vaccine.
Kim
S W
SW
College of Pharmacy, Sungkyunkwan University, Suwon, South Korea.
Choi
I H
IH
Kim
S N
SN
Kim
Y H
YH
Pyo
S N
SN
Rhee
D K
DK
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D003429:Cross Reactions / Q000276:immunology; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics; D016415:Sequence Alignment; D013296:Streptococcus pneumoniae / Q000235:genetics* / Q000276:immunology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins; C063842:dnaK protein, E coli