| Dof proteins are members of a major family of plant transcription factors that share single highly conserved zinc finger, which appear to consist of two major domains: an N-terminal conserved DNA-binding domain and a C-terminal domain for transcriptional regulation. Recent studies have indicated their important control roles in development process of higher plants,including light, phytohormone and defense responses, seed development and germination.Because the genome is relatively complex and relative research on Dof transcription factor has been lagged behind.Previous research indicated that, in the cDNA library of developing seeds from Peanut (Arachis hypogaea L.),eight clones of Dof cDNA have been found,but the founction was unknown. Therefore, peanut Dof gene discovery and functional studies should be carried out urgently.To explore the factors on CTAB method of plant DNA extraction that affect the quantity and quality, seven methods were used to extract genomic DNA from peanut leaves of Arachis hypogaea. The DNA samples obtained were detected by appearance,agarose gel electrophoresis and ultraviolet spectrophotometry. Based on the comparative analysis of yield and quality of the DNA samples by different methods, and considered of all index including yield, protein content and the integrity of DNA molecules, the"phenol-chloroform"method was found to be the most suitable for extraction of genomic DNA from leaves of Arachis hypogaea.According to the cDNA sequence of Dof gene D08 from peanut developing seeds, designed a pair of specific primers and successfully cloned varieties of D08 genes with eight varieties of peanut genomic DNA as templates.By comparing the characteristics of D08 gene between nucleotide sequences and the predicted amino acid sequences, analyzed the relationship between the variety and the sequence,got the possible function of D08 genes.The result showed, the"Zhong Hua 4 Hao"has a obviously difference compairing with other peanut. |
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