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Bulletin of Botanical Research ›› 2017, Vol. 37 ›› Issue (2): 194-199.doi: 10.7525/j.issn.1673-5102.2017.02.006

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Construction and Expression of mCherry Expression Vector Used in Plant

YU Ying, LIU Jia-Xin, ZHOU Mei-Qi, ZHAO Lei-Fei, WANG Chao   

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040
  • Received:2016-11-08 Online:2017-03-15 Published:2017-03-11
  • Supported by:
    National High Technology Research and Development Program 863(2013AA102704)

Abstract: Fluorescent proteins were widely used and played important roles in the biological research. The red fluorescent protein mCherry is valuable to the plant research because of its color and stability. The mCherry gene was inserted into pBI121 plant expression vector, and the pBI121MCS-mCherry vector was constructed. The vector were transferred into onion epidermis by particle bombardment, and the whole onion cells showed red fluorescence under the microscope, which suggested that the vector could express red fluorescent protein in plant cells. The transcription factor BpMYB4 gene was constructed in this vector using double enzyme digestion, and the fusion expression vector pBI121MCS-mCherry-BpMYB4 was obtained and expressed in onion epidermis. Only the nucleus showed red fluorescence, which indicated that the vector can be used to express the fusion protein and used in the subcellular localization accurately. While the fusion expression vector can be constructed simply, and the intermediate vectors are no longer required. The KpnⅠ sites was introduced which can help the fusion of more genes. Therefore, mCherry vector can be used in cell localization of plant genes and screening of transgenic plants. This study would provide materials for genomics research of birch in future.

Key words: mCherry, red fluorescent protein, plant expression vector construction, cell localization

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