論文 - 詳細
| RRC ID | 17428 |
|---|---|
| 著者 | Koyama T, Conlon JM, Iwamuro S. |
| タイトル | Molecular cloning and characterization of cDNAs encoding biosynthetic precursors for the antimicrobial peptides japonicin-1Ja, japonicin-2Ja, and temporin-1Ja in the Japanese brown frog, Rana japonica. |
| ジャーナル | Zoolog Sci |
| Abstract |
Using a combination of reverse-transcription polymerase chain reaction and the 5'- and/or 3'-rapid amplification of cDNA ends, we cloned, from a Japanese brown frog (Rana japonica) skin total RNA preparation, cDNAs encoding biosynthetic precursors for the antimicrobial peptides (AMPs) japonicin-1Ja (FFPIGVFCKIFKTC), japonicin-2Ja (FGLPMLSILPKALCILLKRKC), and temporin-1Ja (ILPLVGNLLNDLL.NH2). These peptides were previously isolated from an extract of R. japonica skin. The present study is the first report to describe the molecular cloning of the cDNA encoding a japonicin-2 family peptide. The nucleotide and deduced amino acid sequence analyses revealed that the hypothetical precursor protein of japonicin-2Ja, as well as japonicin-1Ja and temporin-1Ja, is organized similarly to those of typical amphibian AMP precursors, with a highly conserved signal peptide, a relatively well conserved intervening sequence, and a hypervariable AMP mature region. Antimicrobial assays for synthetic replicates of cyclic and linear japonicin-2Ja revealed that the intramolecular disulfide bond is necessary for activity. A semi-quantitative analysis by real-time RTPCR using TaqMan probes revealed that the relative values of preprojaponicin-2Ja mRNA expression levels in the skin, skeletal muscle of hind leg, kidney, testis, small intestine, and stomach total RNA sample specimens in adult R. japonica were 6.5×10(5), 9.6, 2.0, 1.6, 1.6, and 1.0, respectively. The presence of preprojaponicin-2Ja mRNAs in the cytoplasm of glandular cells in R. japonica dorsal skin glands was demonstrated by means of in situ hybridization using digoxigenin-labeled cRNA probes for the precursor. |
| 巻・号 | 28(5) |
| ページ | 339-47 |
| 公開日 | 2011-5-1 |
| DOI | 10.2108/zsj.28.339 |
| PMID | 21557657 |
| MeSH | Amino Acid Sequence Animals Anti-Infective Agents Base Sequence Cloning, Molecular* DNA, Complementary / genetics* DNA, Complementary / metabolism Gene Expression Regulation / physiology* Molecular Sequence Data RNA, Messenger / genetics RNA, Messenger / metabolism Ranidae / metabolism* |
| IF | 0.843 |
| 引用数 | 6 |
| WOS 分野 | ZOOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 一般微生物 | JCM 2874 JCM 5491 |