Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (7): 1314-1322.doi: 10.3864/j.issn.0578-1752.2013.07.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Characterization of the BnbZIP1 Transcription Factor Gene from Ramie (Boehmeria nivea L.)

 ZHOU  Jing-Hua, JIE  Yu-Cheng, XING  Hu-Cheng, ZHONG  Ying-Li, YU  Wei-Lin   

  1. 1.Institute of Ramie, Hunan Agricultural University, Changsha 410128
    2. Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha 410128
    3. College of Bioscience and Biotechnology, Hunan Agricultural University,        Changsha 410128
  • Received:2012-12-19 Online:2013-04-01 Published:2013-02-04

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Abstract: 【Objective】The objective of this study was to clone the full-length cDNA of BnbZIP1 transcription factor gene from ramie, and the expression pattern and the bioinformatics of the sequence were analyzed, and the prokaryotic expression and the subcellular localization were analyzed.【Method】A full-length cDNA sequence was cloned by RT-PCR and RACE methods based on the unigene48047 in ramie transcriptome sequencing. Then the sequence was analyzed through bioinformatics methods and the expression patterns of BnbZIP1 were analyzed by using Real-time PCR in various tissues and under different stress conditions. A prokaryotic expression vector was constructed and the prokaryotic protein expression was induced with IPTG. Then a fusion expression vector containing EGFP was constructed to observe the subcellular localization. 【Result】The full-length cDNA sequence and the ORF of BnbZIP1 were 2 071 bp and 1 407 bp, which encoded 468 amino acids with predicted pI and molecular weight were 4.95 and 36.81 kD, respectively. Homology comparison analysis showed that the deduced BnbZIP1 amino acid sequence shares a 93% homology with bZIP gene (XP_002307972) in Populus trichocarpa. The relative molecular weight of recombinant protein induced by IPTG was 52 kD, which corresponded to the theoretical value. The results of subcellular localization analysis showed that the BnbZIP1 is located in nucleus. The results of real-time PCR suggested that the BnbZIP1 gene expressed in root, stem, shoot tip and blade, female flowers and male flowers, with the highest expression level in male flowers and the lowest in root. The BnbZIP1 gene was up-regulated by ABA, drought and high salt treatment.【Conclusion】The full-length cDNA sequence of BnbZIP1 from ramie was cloned and it had the typical bZIP transcription factor structural domain in plants, and BnbZIP1 gene responses to ABA, drought and high salt stress, which indicated that the BnbZIP1 genes might play an important role in stress response.

Key words: ramie , transcription factor , bZIP , expression analysis , subcellular localization

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