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Acta Prataculturae Sinica ›› 2012, Vol. 21 ›› Issue (4): 168-174.

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A study on Agrobacterium tumefaciens-mediated transformation of Medicago sativa with the AtCBF1 gene

XU Chun-bo1,2, WANG Yong1, ZHAO Hai-xia2, MI Fu-gui1   

  1. 1.College of Ecology and Environment, Inner Mongolia Agricultural University, Huhhot 010018, China;
    2.Grassland Research Institute, Chinese Academy of Agricultural Sciences, Huhhot 010010, China
  • Received:2011-06-20 Online:2012-04-25 Published:2012-08-20

Abstract: The cold resistance effect of the cold-induced transcription activator CBF1 (C-repeat-binding-factor) gene on Medicago sativa was studied by amplifying an AtCBF1 gene and cloning it by polymerase chain reaction (PCR) from the genomic DNA of Arabidopsis thaliana. The length of the DNA cloned fragment was 642 bp. Compared with the CBF1 sequence in GenBank, the sequence homology rate was 99.84%. On the basis of the CBF1 gene, the plant expression vector pBI121-CBF1 was constructed with the AtCBF1 gene and M. sativa was transformed using Agrobacterium tumefaciens. The transformed plants were selected with kanamycin. An electrophoretic band of 650 bp was obtained by PCR and RT-PCR (reverse transcription polymerase chain reaction). The AtCBF1 gene was expressed in M. sativa. This research provides a good basis for breeding new M. sativa varieties with cold resistance.

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