香菇順式調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：應用順式調控元件探測載體G221構建了一個香菇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,能以轉錄增強活性篩選香菇DNA片段.用這個基因組文庫轉化酵母菌,獲得了一批lacZ陽性轉化子.對其中表達較強的陽性轉化子進行質粒抽提和雙酶切鑒定,篩選到50個香菇順式調控元件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錄相關的特征序列.該研究也探討了利用酵母表達系統克隆香菇順式調控元件的可行性.
關鍵詞：食用菌; 基因組文庫; 轉錄增強活性; 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農重點攻關項目資金資助
作者簡介：胡樂琴:huyueqin@yahoo.com,通訊作者潘迎捷
作者單位：胡樂琴（南京農業大學資環學院,南京,210095）
姚泉洪（上海農科院生物技術研究中心,上海,201106）
陳明杰（上海農科院食用菌研究所,農業部食用菌遺傳育種重點開放實驗室,上海,201106）
熊愛生（上海農科院生物技術研究中心,上海,201106）
潘迎捷（上海農科院食用菌研究所,農業部食用菌遺傳育種重點開放實驗室,上海,201106）
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