Abstract
We have purified and partially characterized a human platelet aggregation factor (Sm-hPAF) from extracellular products of Streptococcus mitis, strain Nm-65 isolated from a patient with Kawasaki disease. Chemical analysis revealed that Sm-hPAF contains first 15 amino-terminal residues were N-DEQGNRPVETENIAR. Based on this partial sequence, sm-hpaf that encodes Sm-hPAF was amplified by PCR. Sequence analysis indicated that sm-hpaf encodes 665 amino acids with 36 residues as signal sequene. Deduced amino acid sequence of sm-hpaf without signal peptides was named Sm-hPAF. It suggested that partial sequence was similar to cytolysin family (Perfuringolysin O, Intermedilysin, and Streptolysin O etc.), but no homology/similarity was seen in N-terminal sequence of Sm-hPAF. It was suggested that Sm-hPAF had new domain except for four domains that were highly conserved among cytolysin family and designated as domain 0. Each domain was amplified and cloned into pQE 30, 31 and 32 vectors. The vectors were transformed into Escherichia coli M15[pREP4] and expressed with IPTG. Aggregation activity of each recombinant product is in progress.
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Ohkuni, H., Watanabe, Y., Todome, Y. et al. Cloning and Domain Expressions of Streptococcus mitis-derived Human Platelet Aggregation Factor (SM-HPAF) Gene in Escherichia Coli. Pediatr Res 53, 169 (2003). https://doi.org/10.1203/00006450-200301000-00091
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DOI: https://doi.org/10.1203/00006450-200301000-00091