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| 香菇顺式调控元件的克隆及其序列分析* | |
| 引用本文： | 胡乐琴,姚泉洪,陈明杰,熊爱生,潘迎捷.香菇顺式调控元件的克隆及其序列分析*[J].菌物学报,2002,21(3):406-411. |
| 作者姓名： | 胡乐琴 姚泉洪 陈明杰 熊爱生 潘迎捷 |
| 作者单位： | 1. 南京农业大学资环学院,南京,210095 2. 上海农科院生物技术研究中心,上海,201106 3. 上海农科院食用菌研究所,农业部食用菌遗传育种重点开放实验室,上海,201106 |
| 基金项目： | 上海科技兴农重点攻关项目资金资助 |
| 摘 要： | 应用顺式调控元件探测载体G221构建了一个香菇Lentinula edodes基因组文库。G221为大肠杆菌-酿酒酵母穿梭载体,含有一个由酵母Cyc1基因基本启动子控制的lacZ标记基因,能以转录增强活性筛选香菇DNA片段。用这个基因组文库转化酵母菌,获得了一批lacZ阳性转化子。对其中表达较强的阳性转化子进行质粒抽提和双酶切鉴定,筛选到50个香菇顺式调控元件DNA片段。对其中部分片段进行了测序,并对其中一个序列进行了序列分析,鉴别了该序列上的几个与转录相关的特征序列。该研究也探讨了利用酵母表达系统克隆香菇顺式调控元件的可行性。 |
| 关 键 词： | 食用菌 基因组文库 转录增强活性 LacZ |
| 文章编号： | 1007-3515(2002)03-0406-0411 |
| 修稿时间： | 2002年3月18日 |
CLONING AND SEQUENCE ANALYSIS OF CIS-ACTING ELEMENT OF LENTINULA EDODES |
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| Authors: | HU Yue-Qin YAO Quan-Hong CHEN Ming-Jie XIONG Ai-Sheng PAN Ying-Jie |
| Institution: | HU Yue-Qin1 YAO Quan-Hong2 CHEN Ming-Jie3 XIONG Ai-Sheng2 PAN Ying-Jie3 |
| Abstract: | After the genomic DNA was fully digested with Sau3AI , the genomic DNA library of Lentinulu edodes was constructed by inserting the digested genomic DNA fragments into the upstream of the yeast Cyc1 gene mini-promoter which control the expression of the lacZ reporter gene in a E.coli-yeast shuttle vector ,G221. By transformation of the plasmid library into Saccharomyces.cerevisiae , 50 positive plasmids containing the cis-acting elements of Lentinula edodes were obtained. One of these cis-acting elements was sequenced and compared with other nucleotide sequences in the database .Several motifs were identified and analysed .This study also demonstrates that cloning of cis-acting element of edible fungi is feasible using the Saccharomyces.cerevisiae lacZ reporter plasmid system. |
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