| 引用本文: | 胡廷章, 杨俊年, 陈再刚, 吴应梅.水稻OsGSTLc启动子的分离和鉴定[J].广西植物,2014,(2):256-262.[点击复制] |
| HU Ting-Zhang, YANG Jun-Nian, CHEN Zai-Gang, WU Ying-Mei.Isolation and identify of the rice OsGSTLc promoter[J].Guihaia,2014,(2):256-262.[点击复制] |
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| 摘要: |
| 半定量RT-PCR分析表明,OsGSTLc在水稻根中的表达受绿磺隆的诱导。从水稻基因组中分离到的OsGSTLc读码框上游2 2171 bp序列,在起始密码ATG上游-86 bp处有CAAT-box,但在CAAT-box与读码框之间没有典型的TATA-box。因此,OsGSTLc启动子是无TATA框启动子。将OsGSTLc启动子5’-端系列缺失后,分别与GUS报告基因融合,获得GSTL2171::GUS、GSTL1761::GUS、GSTL962::GUS和GSTL525::GUS表达载体,利用农杆菌介导转化水稻,获得转基因水稻,均能启动下游GUS报告基因的表达。氯磺隆处理后,转入GSTL2171::GUS、GSTL1761::GUS和GSTL962::GUS的水稻植株根部的GUS活性明显增加。氯磺隆诱导的应答元件在-962～-525 bp的范围内。 |
| 关键词: OsGSTLc 诱导 缺失分析 启动子 水稻 |
| DOI：10.3969/j.issn.1000-3142.2014.02.022 |
| 分类号:Q71 |
| 基金项目: |
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| Isolation and identify of the rice OsGSTLc promoter |
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HU Ting-Zhang*, YANG Jun-Nian, CHEN Zai-Gang, WU Ying-Mei
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School of Life Sciences and Engineering, Chongqing Three Gorges University, Chongqing 404100, China
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| Abstract: |
| The expression of OsGSTLc in rice roots was induced by chlorsulfuron through semi-quantitative RT-PCR analysis demonstrated. A 2 171 bp upstream sequence of the translation start codon ATG of OsGSTLc gene was isolated from the genomic DNA of rice,which contained a putative CAAT-box at position -86 bp upstream of ATG,but there wasn’t TATA-box between the CAAT-box and ATG. Therefore,OsGSTLc promoter was a TATA-less promoter. To define the core promoter sequence,a series of 5’ truncation derivatives of GST2171 were fused to a GUS reporter gene to construct GSTL2171::GUS,GSTL1761::GUS,GSTL962::GUS and GSTL525::GUS,respectively. The four fusion genes were introduced into rice plants by Agrobacterium-mediated transformation,all promoter fragments with 5’-deletion drived successfully the expression of GUS report gene. Quantitative fluorescence assays showed that the GUS activities in the roots of GSTL2171::GUS,GSTL1761::GUS and GSTL962::GUS transgenic rice seedlings were upregulated by chlorsulfuron. The transcriptional activation element of chlorsulfuron may be located between positions -962 and -525.The expression of OsGSTLc in rice roots was induced by chlorsulfuron through semi-quantitative RT-PCR analysis demonstrated. A 2 171 bp upstream sequence of the translation start codon ATG of OsGSTLc gene was isolated from the genomic DNA of rice,which contained a putative CAAT-box at position -86 bp upstream of ATG,but there wasn’t TATA-box between the CAAT-box and ATG. Therefore,OsGSTLc promoter was a TATA-less promoter. To define the core promoter sequence,a series of 5’ truncation derivatives of GST2171 were fused to a GUS reporter gene to construct GSTL2171::GUS,GSTL1761::GUS,GSTL962::GUS and GSTL525::GUS,respectively. The four fusion genes were introduced into rice plants by Agrobacterium-mediated transformation,all promoter fragments with 5’-deletion drived successfully the expression of GUS report gene. Quantitative fluorescence assays showed that the GUS activities in the roots of GSTL2171::GUS,GSTL1761::GUS and GSTL962::GUS transgenic rice seedlings were upregulated by chlorsulfuron. The transcriptional activation element of chlorsulfuron may be located between positions -962 and -525. |
| Key words: OsGSTLc induction deletion analysis promoter rice |
