TY - JOUR
T1 - Cloning and sequencing of BES-1 gene encoding the immunogenic antigen of Streptococcus sanguis KTH-1 isolated from the patients with Behcet's disease
AU - Yoshikawa, Koji
AU - Kotake, Satoshi
AU - Kubota, Toru
AU - Kimura, Koichi
AU - Isogai, Emiko
AU - Fujii, Nobuhiro
N1 - Funding Information:
This study was supported in part by grant 06671746 from the Ministry of Education, Science, and Culture in Japan and the Research Committee of Beh~et's disease in Japan.
PY - 1998
Y1 - 1998
N2 - In order to analyze the immunopathologic mechanisms of Behcet's disease, the gene (bes-1) encoding a streptococcal antigen correlated with the disease was cloned and sequenced, and protein produced by this clone was identified by Western immunoblotting using serum antibody from the patient. Cellular DNA of Streptococcus (S.) sanguis serotype KTH-1 (uncommon serotype 1, strain 113-20) from the patient was extracted and digested with EcoRI. The digested fragments were cloned into the cloning vector λgt11, and then the resulting DNA library was immunoscreened using the patient's serum antibody to serotype KTH-1. The immunopositive clone of the 1.5 kbp fragment was subcloned into pUC 118 plasmid (pU8BeS1-1) and sequenced. The sequence showed that the 3'-terminal half side region of this insert contained 962bp of open-reading frame (ORF) discontinued at the EcoRI restriction site, and the stop codon was not found. The nucleotide sequence of the remaining additional 3'-terminal region of this gene encoding whole BES-1 was determined by genome walking. The whole ORF of bes-1 consisted of 849 amino acid residues with a calculated molecular mass of 95 kDa. The residues in a portion of the amino acid sequence showed a 60% correspondence to those of the human intraocular peptide Brn-3b.
AB - In order to analyze the immunopathologic mechanisms of Behcet's disease, the gene (bes-1) encoding a streptococcal antigen correlated with the disease was cloned and sequenced, and protein produced by this clone was identified by Western immunoblotting using serum antibody from the patient. Cellular DNA of Streptococcus (S.) sanguis serotype KTH-1 (uncommon serotype 1, strain 113-20) from the patient was extracted and digested with EcoRI. The digested fragments were cloned into the cloning vector λgt11, and then the resulting DNA library was immunoscreened using the patient's serum antibody to serotype KTH-1. The immunopositive clone of the 1.5 kbp fragment was subcloned into pUC 118 plasmid (pU8BeS1-1) and sequenced. The sequence showed that the 3'-terminal half side region of this insert contained 962bp of open-reading frame (ORF) discontinued at the EcoRI restriction site, and the stop codon was not found. The nucleotide sequence of the remaining additional 3'-terminal region of this gene encoding whole BES-1 was determined by genome walking. The whole ORF of bes-1 consisted of 849 amino acid residues with a calculated molecular mass of 95 kDa. The residues in a portion of the amino acid sequence showed a 60% correspondence to those of the human intraocular peptide Brn-3b.
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U2 - 10.1016/S0934-8840(98)80184-9
DO - 10.1016/S0934-8840(98)80184-9
M3 - Article
C2 - 9638874
AN - SCOPUS:0032077952
VL - 287
SP - 449
EP - 460
JO - International Journal of Medical Microbiology
JF - International Journal of Medical Microbiology
SN - 1438-4221
IS - 4
ER -