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Plant cytochrome P450s are involved in the biosynthesis of various classes of secondary metabolites. To elucidate the biosynthesis of zerumbone, a sesquiterpenoid with multiple potential anticancer properties, a family of P450 genes expressed in rhizomes of Zingiber zerumbet Smith, were cloned using a PCR-based cloning strategy. After functional expression in yeast, one of these P450s was found to convert α-humulene into 8-hydroxy-α-humulene, a proposed intermediate of zerumbone biosynthesis. This P450 has been designated CYP71BA1, a new member of the CYP71 family. CYP71BA1 transcripts were detected almost exclusively in rhizomes and showed a similar expression pattern to ZSS1 transcripts during rhizome development. Coexpression of a gene cluster encoding four enzymes of the mevalonate pathway with CYP71BA1 and ZSS1 in Escherichia coli leads to the production of 8-hydroxy-α-humulene in the presence of mevalonate, suggesting the possibility of microbial production of this zerumbone intermediate from a relatively simple carbon source by metabolic engineering. [ABSTRACT FROM AUTHOR]

Our previous study demonstrated that among the various T-cell receptor Vb gene segments, T-cell receptor Vb6 gene-bearing T cells are predominant in periodontitis lesions. To explore the clonality of T cells expressing Vb6 T-cell receptor, which is determined by the heterogeneity of the complementarity-determining region 3(CDR3), Vb6-Cb polymerase chain reaction products derived from gingival tissue and peripheral blood of periodontitis patients were analyzed by single-strand conformation polymorphism and nucleotide sequence. Both techniques provided quite similar results in terms of the number of accumulated clones in which clonal accumulation of T-cell receptor Vb6-bearing T cells was found in most periodontitis lesions but not in peripheral blood. From the analysis of deduced amino acid sequences, a possible conserved amino acid residue in the CDR3 region of the periodontitis lesion-derived sequences was found in most patients. This conserved amino acid motif however, was not identified. Furthermore, skewed Jb gene segment usage was observed for expanded clones from periodontitis lesions. Our results suggest that T cells infiltrating periodontitis lesions are responding to a restricted number of antigens or epitopes. [ABSTRACT FROM AUTHOR]