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| Functional Analysis of the ZAG2 Promoter from Maize in Transgenic Tobaccos |
| LU Min-hui, WANG Guo-ying, MENG Zheng, WANG Jian-hua |
1.Plant Genetic and Breeding, China Agricultural University, Beijing 100193, P.R.China
2.Institute of Crop Sciecnes, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
3.Center for Signal Transduction and Metabolomics, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, P.R.China |
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摘要 The function of the 3 040 bp sequence at the upstream translation starting site (ATG) of the ZAG2 gene, isolated from the maize genome, was studied. The sequence analysis showed that the sequence contained a typical class C MADS-box gene regulatory element. The 5´ UTR region of the gene contains a 1 299-bp intron that might have important regulatory functions. To study the sequence function, deletion derivatives of promoter-reporter (uidA) gene fusions were generated and transformed into tobaccos. The GUS staining and fluorescence quantification results showed that the GUS activity was detected only in the third and fourth whorl floral organs of the transgenic tobaccos under driving the promoter including the first intron, while detected in all the organs and was stronger under driving the promoter without the first intron. However, the GUS activity was just detected in one whorl of the fourth or third floral organs under driving of the 35S promoter. These results suggested that the first intron of the ZAG2 gene contains functional regulatory elements, which turned out to be important for gene expression in the heterologous systems. Moreover, the GUS activity was decreased when the reporter gene driven by the promoters with 5´-deletions, respectively, from -1 606 to -951 and -951 to -426 nts, which indicates that positive regulatory elements are present in these two sequence stretches.
Abstract The function of the 3 040 bp sequence at the upstream translation starting site (ATG) of the ZAG2 gene, isolated from the maize genome, was studied. The sequence analysis showed that the sequence contained a typical class C MADS-box gene regulatory element. The 5´ UTR region of the gene contains a 1 299-bp intron that might have important regulatory functions. To study the sequence function, deletion derivatives of promoter-reporter (uidA) gene fusions were generated and transformed into tobaccos. The GUS staining and fluorescence quantification results showed that the GUS activity was detected only in the third and fourth whorl floral organs of the transgenic tobaccos under driving the promoter including the first intron, while detected in all the organs and was stronger under driving the promoter without the first intron. However, the GUS activity was just detected in one whorl of the fourth or third floral organs under driving of the 35S promoter. These results suggested that the first intron of the ZAG2 gene contains functional regulatory elements, which turned out to be important for gene expression in the heterologous systems. Moreover, the GUS activity was decreased when the reporter gene driven by the promoters with 5´-deletions, respectively, from -1 606 to -951 and -951 to -426 nts, which indicates that positive regulatory elements are present in these two sequence stretches.
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Received: 03 May 2011
Accepted:
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| Fund: This program was financially supported by the National Major Project for Transgenic Organism Breeding, China (2011ZX08003-001). |
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Corresponding Authors:
Correspondence WANG Jian-hua, Tel/Fax: +86-10-62732263, E-mail: wangjh63@cau.
edu.cn; MENG Zheng, Tel/Fax: +86-10-62836556, E-mail: zhmeng@ibcas.ac.cn
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| About author: LU Min-hui, Tel/Fax: +86-10-62733410, E-mail: luminhui-2001@163.com |
Cite this article:
LU Min-hui, WANG Guo-ying, MENG Zheng, WANG Jian-hua.
2012.
Functional Analysis of the ZAG2 Promoter from Maize in Transgenic Tobaccos. Journal of Integrative Agriculture, 12(8): 1266-1273.
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