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)控元件探測載體G221構(gòu)建了一個香菇Lentinula edodes基因組文庫.G221為大腸桿菌-釀酒酵母穿梭載體,含有一個由酵母Cyc1基因基本啟動子控制的lacZ標記基因,能以轉(zhuǎn)錄增強活性篩選香菇DNA片段.用這個基因組文庫轉(zhuǎn)化酵母菌,獲得了一批lacZ陽性轉(zhuǎn)化子.對其中表達較強的陽性轉(zhuǎn)化子進行質(zhì)粒抽提和雙酶切鑒定,篩選到50個香菇順式調(diào)控元件DNA片段.對其中部分片段進行了測序,并對其中一個序列進行了序列分析,鑒別了該序列上的幾個與轉(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; 轉(zhuǎn)錄增強活性; LacZ

分類號：Q939.96 　文獻標識碼：A

文章編號：1007-3515(2002)03-0406-0411

基金項目：本研究由上?？萍寂d農(nóng)重點攻關(guān)項目資金資助

作者簡介：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)大學資環(huán)學院,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開放實驗室,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開放實驗室,上海,201106）　

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