DOF转录因子AtDof1.7 RNA干扰载体的构建及拟南芥的遗传转化

doi:10.3724/SP.J.1006.2011.01196

Abstract

Abstract: The Dof (DNA binding with one finger) transcription factorsare members of a family of plant-specific transcription factors that have a highly conserved DNA-binding domain, namely Dof domain which contains a single C2C2-type zinc-finger-like motif and specifically recognizes an (A/T)AAAG sequence as the recognition core. It suggests that the Dof transcription factors play diverse roles in specific biological processes in plants. Members of this protein family in plants are found to be involved in the gene regulation of many processes, but the roles in fatty acid metabolism are rarely reported. To investigate whether AtDof1.7 Dof transcription factor can regulate fatty acid metabolism, on the basis of the sequence of the gene (GenBank accession No. AT1G51700), we designed the specific primer containing different enzyme sites. With the template of Arabidopsis thaliana DNA, the AtDof1.7 gene fragment was isolated, which wasinserted into the expression vector by forward and reverse ways respectively. The RNA interference vector of pADOF1 containing AtDof1.7 gene fragment was constructed. By floral-dip method, pADOF1 was successfully transformed into wide-type Arabidopsis thaliana. Glyphosate resistance screening and PCR detection showed that five positive transgenic plants were obtained. The result of RT-PCR showed that transgenic plants had lower expression level of AtDof1.7 gene than the wild type. Fatty acid content was analyzed by gas chromatography which showed that the content of oleic acid increased and the content of linolenic acid decreased drastically in each transgenic plant compared with wide-type plants. These results indicated that AtDof1.7 transcription factor has certain relation with fatty acid metabolic pathway in Arabidopsis thaliana seed which provides a good foundation for further study on the function of AtDof1.7 transcription factor in fatty acid metabolic regulation.

Key words: DOF transcription factors, AtDof1.7, Arabidopsis thaliana, RNA interference vector, Fatty acid metabolic pathway

YIN Meng-Zhi, GUAN Mei, XIAO Gang, LI Quan, GUAN Chun-Yun. RNAi Vector Construction of AtDof1.7 Transcription Factors and Genetic Transformation into Arabidopsis thaliana[J].Acta Agron Sin, 2011, 37(07): 1196-1204.

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RNAi Vector Construction of AtDof1.7 Transcription Factors and Genetic Transformation into Arabidopsis thaliana

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