臺灣博碩士論文加值系統

隨著分子生物學技術的迅速發展，真菌的分類學己經從表型形態分類進入了基因分子分類的新時代。本實驗中利用聚合酶連鎖反應（Polymerase Chain Reaction）技術，以靈芝漆酶（laccase）基因之共同序列（consensus sequence）及核糖體DNA內轉錄間隔區序列 （ribosomal DNA internal transcribed spacer region sequences ; ITS）為標的，分別對15株靈芝(4株標準菌株及11株野外分離株)及3株外群菌屬，PCR增幅出ITS1-5.8S-ITS2序列 595~915 bp及Laccase 序列1401~1651 bp之片段，並分別以TA-技術選殖後進行DNA定序。所得的DNA序列以NCBI網站資料庫進行相似性序列比對分析，用以確認分子鑑定與傳統型態分類命名的異同。並分別將DNA序列以CLUSTAL程式進行親緣關係比對分析，以建立演化之親緣關係樹。

With the rapid development of molecular biology techniques, the taxonomy of fungi has been transformed from by morphological phenotype into a new era of molecular classification. In this study, we use of polymerase chain reaction (PCR) technology to amplify both the consensus sequence of the laccase gene and the ribosomal DNA internal transcribed spacer sequences (ITS) of 15 Ganoderma strains (including four BCRC standard strains and 11 of wild type isolates) and three exceptionally genus strains. The specific 595~915 bp of PCR product of TS1-ITS2 fragement and 1401~1651 bp of laccase gene fragments have been amplified, then cloning by TA- strategy for DNA sequencing. Each DNA sequence has been applied into NCBI database for sequence alignment by similarity analysis to compare the naming of molecular identification and traditional morphological classification. And, respectively, alignment by CLUSTAL program to establish the phylogenetic tree.

第一章 前言…………………………………………………1
第二章 前人研究……………………………………………2
第三章 材料與方法…………………………………………4
第四章 結果………………………………………………..10
第五章 討論 ………………………………………………24
參考文獻……………………………………………………26
附錄1 ITS序列分析表………………………………….….29
附錄2 Laccase序列分析表…………………………………53

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