1995 Fiscal Year Final Research Report Summary
Identification of a Pathogenic Streptococcal Antigen in Behcet's disease
| Project/Area Number |
06671746
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Ophthalmology
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| Research Institution | Hokkaido University |
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Principal Investigator |
KOTAKE Satoshi Hokkaido University, University Hospital, Lecturer, 医学部附属病院, 講師 (00186694)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIKAWA Koji Hokkaido University, School of Medicine, Instructor, 医学部, 助手 (00250431)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Behcet's disease / Streptococcus / Uveities / Cloning |
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| Research Abstract |
To analyze immunopathologic mechanisms of Behcet's disease, we cloned and sequenced the gene encoding a streptococcal antigen correlated with this disease. Cellular DNA of Streptococcus (S.) sanguis KTH-1 strain was extracted and digested with EcoRI.The fragments were cloned by using bacteriophage lgtl 1 and Escherichia coli. DNA library of Eco RI fragments was plated and immunoscreened with the serum antibody of a patient with Behcet's disease, who showed a high antibody value to KTH-1 strain. The DNA fragment isolated from immunopositive clones consisted of about 1.5kbp, and then was subcloned into pUC118 plasmid. The nucleotide sequnce of this clone reveled thant the 3'-terminal region of the insert contained a 950bp opne reading frame (ORF) discontinued by digestion with EcoRI.The molecular weight of this region was equal to that of the streptococcal antigen which responsed to serum antibody of the same patient by means of Western blotting. In an attempt to decide the nucleotide sequence of the 3'-terminal region of this ORF,the polymerase chain reaction was employed after ligation of appropriate restriction fragments and cassettes. The entire ORF was starting at ATG (position 603) which was proceeded by Shine-Dalgarno ribosome binding site, and was capable of coding a peptide 2,550 amino acid residues with a calculated molecular mass of about 94,800Da. The amino acid sequence is homologous to that of Brn-3b with 60% identical residues.
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