| 香菇ras和gpd启动子的克隆与功能鉴定 |
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| 引用本文： | 于晓玲,林俊芳,郭丽琼,王树昌.香菇ras和gpd启动子的克隆与功能鉴定[J].食用菌学报,2005,12(3):15-20. |
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| 作者姓名： | 于晓玲 林俊芳 郭丽琼 王树昌 |
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| 作者单位： | 1. 中国热带农业科学院热带作物生物技术国家重点实验室,热带生物技术研究所,海口,571101
2. 中国热带农业科学院热带作物生物技术国家重点实验室,热带生物技术研究所,海口,571101;华南农业大学食品学院生物工程系,广州,510640 |
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| 基金项目： | 国家自然科学基金资助项目“抗冷冻蛋白基因遗传转化草菇的研究”（编号：39960050）,“草菇冷冻胁迫诱导表达基因及其启动子的克隆分离”（编号：30060054）,“食用菌外源基因表达系统的建立”（编号：30371000）和广东省自然科学基金资助项目“食用菌外源基因表达系统的建立”（编号：032239）的部分研究内容 |
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| 摘 要： | 以香菇基因组DNA为模板,利用PCR技术克隆了香菇1个ras启动子Lras,2个gpd启动子Lgpd1和Lgpd2.这3个启动子的大小分别为715bp、1056bp和611bp,与已报道的ras和gpd启动子的同源性很强.对这3个启动子的分析结果表明,它们都含有丰富的启动子顺式作用元件.将这些启动子片段分别与GUS基因连接构建了表达载体,并将这些表达载体导入草菇菌丝体中,检测其活性,最终显示3个片段都有启动GUS基因表达的能力,其中Lgpd1启动子活性最强,Lras启动子次之,Lgpd2最弱.
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| 关 键 词： | 香菇 ras启动子 gpd启动子 原生质体转化 GUS基因 |
| 文章编号： | 1005-9873（2005）03-0015-06 |
| 收稿时间： | 2005-03-31 |
| 修稿时间： | 2005-05-25 |
Cloning and Characterization of ras and gpd Promoters from Lentinus edodes |
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| YU Xiao-ling,Lin Jun-fang,Guo Li-qiong,Wang Shu-chang.Cloning and Characterization of ras and gpd Promoters from Lentinus edodes[J].Acta Edulis Fungi,2005,12(3):15-20. |
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| Authors: | YU Xiao-ling Lin Jun-fang Guo Li-qiong Wang Shu-chang |
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| Institution: | 1. State Key Laboratory of Tropical Crop Biotechnology, The Chinese Academy of Tropical Agricultural Sciences and Institute of Tropical Biological Sciences, Haikou 571101, China; 2 .Department of Bioengineering, College of Focd Sciences, South China Agricultural University, Guangzhou 510640 ,China |
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| Abstract: | The promoter fragments of Lras, Lgpdl and Lgpd2 were isolated from genomic DNA of Lentinus edodes by PCR amplification. DNA sequencing results showed that these promoter fragments have 715bp, 1056bp and 611bp respectively. Blastn analysis results revealed that Lras, Lgpdl and Lgpd2 have high homologous with reported promoters, ras and gpd . Analysis results showed that they all contain many important cis-acting regulatory elements. Fragments of Lras, Lgpdl and Lgpd2 were constructed respectively with pCAMBIA1301 which 35S promoter was deleted. Then the new expression plasmids containing GUS reporter gene were induced into Volvariella volvacea by protoplast electroporation. GUS histochemistry assay results showed that Lgpdl had the strongest promoter activity, Lras had stronger activity and Lgpd2 had slight activity. |
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| Keywords: | Lentinus Modes ras promoter gpd promoter Protoplast transformation GUS |
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