Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/22143
標題: 立枯絲核菌(Rhizoctonia solani K hn) AG-4融合群中質體DNA限 制▇圖譜及選殖之研究
Mapping and cloning of linear plasmid DNA in Rhizoctonia solani K hn AG-4
作者: 李建勳
Lee, Chien-shiun
關鍵字: Rhizoctonia solani
立枯絲核菌
linear plasmid
mapping
cloning
線形質體
限制▇圖譜
選殖
出版社: 植物學系
摘要: 質體 pCHU341是存在於Rhizoctonia solani (strain CHU341) 中的直線 型分子,分子量約 2.6 kb 。本研究目的乃分析 pCHU341之限制▇圖譜及 利用 pCHU341限制▇片段構築一個新轉形載體。質體 pCHU341限制▇切位 ,乃以包含質體之總體 DNA限制▇切位做初步判斷。正確的質體限制▇切 位則用限制▇(AvaI 、BamHI 、BglII、EcoRI 、 HindIII、SacI、 SphI 及 SmaI) 切割純化的質體得到。利用質體限制▇切位並配合分殖技術( subcloning)可製作質體 pCHU341之限制▇圖譜。 pCHU341經限制▇切割 所得之片段可利用選殖作用(cloning)在Escherichia coli中進行大量複 製。選殖片段可被黏接於轉形載體 pUCH1,此新構築的真菌轉形載體有助 於轉形實驗及其他有關質體 pCHU341 功能之研究。
Plasmid pCHU341 is a linear element of Rhizoctonia solani CHU341 the molecular weight of 2.6 Kb . The aim of this study was tried to analyze the restriction map of plasmid pCHU341 and construct a new transformation vector bearing a restriction fragment from pCHU341 . The cut sites of plasmid pCHU314 were deduced roughly from the restriction fragments of total DNA containing plasmid digested with various enzymes . The cut sites of plasmid were deduced from restriction fragments of purified pCHU341 digested with restriction enzymes ( AvaI、 BamHI、 BglII、 EcoRI、 HindIII、 SacI、 SphI and SmaI ) . The restriction map of pCHU341 was made by cut sites of plasmid associated with subcloning technique . The restriction fragments of pCHU341 can be subcloned by a cloning vector in Escherichia coli . The cloned restriction fragments were fused to a fungal transformation vector pUCH1 . The newly constructed vector derived from pUCH1 bearing the restriction fragment of pCHU341 is useful for transformation experiments and another studies about the function of pCHU341 .
URI: http://hdl.handle.net/11455/22143
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