楊國良; 李麗娜; 陸師義; 艾福彪; 楊秀琴
河北大學生物技術中心; 河北大學生物技術中心; 河北大學生物技術中心; 河北大學生物技術中心; 河北大學生物技術中心 保定 071002 ; 保定 071002 ; 保定 071002 ; 保定 071002 ; 保定 071002

【中文摘要】 pDL1是能夠轉化細菌和真菌的雙功能質粒。將pDL1與糙皮側耳(Pleurotus ostreatus)的DNA片段連接,然后轉化玉米黑粉菌(Ustilago maydis)的原生質體。通過新霉素選擇平皿的篩選,轉化率達到800轉化子/μgDNA,建立了糙皮側耳的基因文庫。隨機取15個轉化子進行點漬法(Dotting blotting)檢驗,結果均呈陽性。此結果表明,抗性克隆不是源于玉米黑粉菌敏感細胞的回復突變。薩慎法(Southern blotting)檢驗顯示,本研究在玉米黑粉菌細胞中克隆糙皮側耳纖維二糖水解酶基因是成功的。基因表達試驗表明,玉米黑粉菌克隆株具有糙皮側耳纖維二糖水解酶的活性。

【英文摘要】 pDL1 is a kind of bifunctional plasmid which can transform bacteria and fungi. And pDLl transformed the protoplasts of Ustilago maydis after being ligated with DNA fragments of Pleurotus ostreatus. Through the screening of neomycin selective plates, the transformation ratio reached 800 transformants per microgram of DNA, and the gene library of Pleurotus ostreatus was established. 15 randomly selected transformants expressed positive by dotting blotting test. The result showed that the resistant clones were not originated from back mutation of sensitive cell of Ustilago maydis. And Southern blotting test revealed that studies were successful for cloning the cellobiosehydrolase gene of Pleurotus ostreatus in the cells of Ustilago maydis. While the gene expression test showed that the cloned strains of Ustilago maydis expressed the enzyme activity of cellobiosehydrolase of Pleurotus ostreatus.

【中文關鍵詞】 糙皮側耳; 玉米黑粉菌; 纖維二糖水解酶; 基因克隆
【英文關鍵詞】 Pleurotus ostreatus; Ustilago maydis; Cellobiosehydrolase; Gene cloning
【基金】瑞典自然科學基金;; 河北省自然科學基金

【文獻出處】 食用菌學報,Acta Edulis Fungi,編輯部郵箱,1998年01期 【DOI】CNKI:SUN:SYJB.0.1998-01-000