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題 名 | 植物菌質體之優勢免疫膜蛋白基因imp,idpA及amp之選殖與應用=Cloning and Application of Phytoplasmal Immunodominant Membrane Protein Genes imp, idpA and amp |
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作 者 | 程諭揚; 林長平; | 書刊名 | 植物病理學會刊 |
卷 期 | 19:1 2010.03[民99.03] |
頁 次 | 頁9-18 |
分類號 | 433.4 |
關鍵詞 | 優勢免疫膜蛋白; 聚合酵素連鎖反應; imp基因; idpA基因; amp基因; 反轉錄聚合酵素連鎖反應; Immunodominant membrane protein; Polymerase chain reaction; Imp gene; IdpA gene; Amp gene; RT-PCR; |
語 文 | 中文(Chinese) |
中文摘要 | 植物菌質體之研究顯示優勢免疫膜蛋白為植物菌質體細胞膜之主要蛋白。優勢免疫膜蛋白依據其嵌入於細胞膜上之構型,可區分為三個類型,分別為第一型優勢免疫膜蛋白 Imp (immunodominant membrane protein)、第二型優勢免疫膜蛋白 IdpA (immunodominant membrane protein A) 以及第三型優勢免疫膜蛋白 Amp (antigenic membrane protein)。本研究利用聚合酵素連鎖反應 (PCR) 並配合基因選殖技術,獲得花生簇葉病菌質體 imp 基因之全長序列、台灣梨衰弱病第二群菌質體 imp 基因之部分序列、聖誕紅叢枝誘導性植物菌質體 imp 基因及 idpA 基因之全長序列、日日春葉片黃化病菌質體 imp 基因及 amp 基因之全長序列、台灣泡桐簇葉病菌質體之 imp 基因及 amp 基因之全長序列。比對序列發現屬於同一 16Sr group 中之植物菌質體,彼此間同一型的優勢免疫膜蛋白基因序列具有差異性之區域,可據以區分不同 16Sr group 或 subgroup 之植物菌質體。針對台灣目前僅有同屬於 16SrI 群之日日春葉片黃化病菌質體及台灣泡桐簇葉病菌質體,根據第三型優勢免疫膜蛋白基因 amp 序列間具差異性之區域,設計出對兩菌質體專一性的 PCR 引子對 ampf5/Amp1R 及 Amp1F/ampr4, PCR 可成功區分此二親緣關係相近之植物菌質體。此外,反轉錄聚合酵素連鎖反應 (RT-PCR) 之結果顯示:日日春葉片黃化病菌質體 imp 及 amp 基因以及花生簇葉病菌質體第一型 imp 基因均可於植物菌質體中轉錄出mRNA。 |
英文摘要 | Previous studies have shown that immunodominant membrane protein constitute a major portion of cellular membrane proteins in most phytoplasmas. Genes encoding immunodominant membrane proteins have been identified and sequenced from several taxonomic groups of phytoplasmas. Based on their topologies in the membrane, the phytoplasma immunodominant membrane proteins can be classified into three distinct types: type 1 immunodominant membrane protein Imp, type 2 immunodominant membrane protein IdpA, and type 3 immunodominant membrane protein Amp. No amino acid similarity was revealed among different types of immunodominant membrane proteins. The sequence identity of immunodominant membrane protein genes between phytoplamas was lower than those of their upstream or downstream genes or non-coding regions. In this study, cloning and sequencing of these three types of immunodominant membrane protein genes were performed for three groups of phytoplasmas, including imp genes from the pear decline phytoplasma (PDTWII phytoplasma) and peanut witches'-broom (PnWB) phytoplasma of 16SrII group, idpA gene from poinsettia branch-inducing (PoiBI) phytoplasma of 16SrIII group , and amp genes from periwinkle leaf yellowing (PLY) phytoplasma and paulownia witches'-broom (PaWB-Taiwan) phytoplasma of 16SrI group. Nucleotide sequences of imp, idpA, or amp genes were different among phytoplasmas of the same group. The distinct immunodominant membrane protein gene sequences can be used for differentiating members within the same groups or subgroups of various phytoplasmas. Based on the distinct regions of amp gene PLY phytoplasma and PaWB-Taiwan phytoplasma, PCR primer pairs ampf5/Amp1R and Amp1F/ampr4 specific for identification and classification of the two phytoplasmas were successfully developed. RT-PCR showed that both the imp, and amp genes of PLY phytoplasma and the imp gene of PnWB phytoplasma were transcribed in PLY phytoplasma and PnWB phytoplasma, respectively. |
本系統中英文摘要資訊取自各篇刊載內容。