香菇順式調(diào)控元件的克隆及其序列分析

CLONING AND SEQUENCE ANALYSIS OF CIS-ACTING ELEMENT OF LENTINULA EDODES

胡樂琴　姚泉洪　陳明杰　熊愛生　潘迎捷　

摘　要：應(yīng)用順式調(diào)控元件探測(cè)載體G221構(gòu)建了一個(gè)香菇Lentinula edodes基因組文庫(kù).G221為大腸桿菌-釀酒酵母穿梭載體,含有一個(gè)由酵母Cyc1基因基本啟動(dòng)子控制的lacZ標(biāo)記基因,能以轉(zhuǎn)錄增強(qiáng)活性篩選香菇DNA片段.用這個(gè)基因組文庫(kù)轉(zhuǎn)化酵母菌,獲得了一批lacZ陽(yáng)性轉(zhuǎn)化子.對(duì)其中表達(dá)較強(qiáng)的陽(yáng)性轉(zhuǎn)化子進(jìn)行質(zhì)粒抽提和雙酶切鑒定,篩選到50個(gè)香菇順式調(diào)控元件DNA片段.對(duì)其中部分片段進(jìn)行了測(cè)序,并對(duì)其中一個(gè)序列進(jìn)行了序列分析,鑒別了該序列上的幾個(gè)與轉(zhuǎn)錄相關(guān)的特征序列.該研究也探討了利用酵母表達(dá)系統(tǒng)克隆香菇順式調(diào)控元件的可行性.

關(guān)鍵詞：食用菌; 基因組文庫(kù); 轉(zhuǎn)錄增強(qiáng)活性; LacZ

分類號(hào)：Q939.96 　文獻(xiàn)標(biāo)識(shí)碼：A

文章編號(hào)：1007-3515(2002)03-0406-0411

基金項(xiàng)目：本研究由上海科技興農(nóng)重點(diǎn)攻關(guān)項(xiàng)目資金資助

作者簡(jiǎn)介：胡樂琴:huyueqin@yahoo.com,通訊作者潘迎捷

作者單位：胡樂琴（南京農(nóng)業(yè)大學(xué)資環(huán)學(xué)院,南京,210095）　

　　　　　姚泉洪（上海農(nóng)科院生物技術(shù)研究中心,上海,201106）　

　　　　　陳明杰（上海農(nóng)科院食用菌研究所,農(nóng)業(yè)部食用菌遺傳育種重點(diǎn)開放實(shí)驗(yàn)室,上海,201106）　

　　　　　熊愛生（上海農(nóng)科院生物技術(shù)研究中心,上海,201106）　

　　　　　潘迎捷（上海農(nóng)科院食用菌研究所,農(nóng)業(yè)部食用菌遺傳育種重點(diǎn)開放實(shí)驗(yàn)室,上海,201106）　

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