| Peanut is one of the important oil and economic crops of the world,which blossoms overgroundly while fruits undergroundly. The status of floral organs development is highly associated with the yield of peanut seeds. MADS-box genes are a gene family which distribute all over the plant genome, and are the key gene of regulating plant growth and development by encoding series of transcription factors of significant biological functions. MADS-box genes play an important role in plants, especially in floral meristem, floral organs and various vegetative organs. Taking peanut variety, Minhua 6, as research materials of this study , we designed degenerate primers according to the conserved sequence of MADS-box genes, then isolated and cloned peanut's MADS-box gene seguence by homology cloning approach. We also analysised the sequence characteristics and the evolutionary relationship, which can be further used to improve the yield of peanut by plant genetic engineering in the future.On the basis of two full-length peanut MADS-box genes our reseach team have acquired and logined into the Genebank previously, as well as homologous sequence of other leguminous plants,we designed 3 forward primers and 3 reverse primers according to the conserved sequences of MADS-box, and matched 5 pairs primer to amplify the fragment by the template of mixed cDNA from different organizations. We obtained 5 targeted fragments, the length were 520bp, 644bp, 433bp, 557bp and 448bp respectively, which were as to our expectation; and then, these fragments were clone into pGEM-T vector, picked up the clones and identified by the PCR and restricted enzyme digest. The clones were analysis by DNA sequencing. At last we blast the sequence of the 26 fragments, and suggesting that all of the fragments were contain the conserved K-box and partial MADS-box of MADS-box. Cluster analysis of the 26 sequences by software Cluster X 2.0 suggesting that those sequences were clustered into different clusters. |
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