楊國良; 李麗娜; 陸師義; 艾福彪; 楊秀琴
河北大學(xué)生物技術(shù)中心; 河北大學(xué)生物技術(shù)中心; 河北大學(xué)生物技術(shù)中心; 河北大學(xué)生物技術(shù)中心; 河北大學(xué)生物技術(shù)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(zhuǎn)化細(xì)菌和真菌的雙功能質(zhì)粒。將pDL1與糙皮側(cè)耳(Pleurotus ostreatus)的DNA片段連接,然后轉(zhuǎn)化玉米黑粉菌(Ustilago maydis)的原生質(zhì)體。通過新霉素選擇平皿的篩選,轉(zhuǎn)化率達(dá)到800轉(zhuǎn)化子/μgDNA,建立了糙皮側(cè)耳的基因文庫。隨機(jī)取15個轉(zhuǎn)化子進(jìn)行點(diǎn)漬法(Dotting blotting)檢驗,結(jié)果均呈陽性。此結(jié)果表明,抗性克隆不是源于玉米黑粉菌敏感細(xì)胞的回復(fù)突變。薩慎法(Southern blotting)檢驗顯示,本研究在玉米黑粉菌細(xì)胞中克隆糙皮側(cè)耳纖維二糖水解酶基因是成功的。基因表達(dá)試驗表明,玉米黑粉菌克隆株具有糙皮側(cè)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(guān)鍵詞】 糙皮側(cè)耳; 玉米黑粉菌; 纖維二糖水解酶; 基因克隆
【英文關(guān)鍵詞】 Pleurotus ostreatus; Ustilago maydis; Cellobiosehydrolase; Gene cloning
【基金】瑞典自然科學(xué)基金;; 河北省自然科學(xué)基金

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