Chinese Bulletin of Botany ›› 2015, Vol. 50 ›› Issue (3): 354-362.DOI: 10.3724/SP.J.1259.2015.00354

• EXPERIMENTAL COMMUNICATIONS • Previous Articles     Next Articles

Studies of the Transient Expression and Transformation of Cloned Thermostable α-Amylase Genes from Bacillus licheniformis in Tobacco and Arabidopsis

Xiao Li1, Haiyan Sun1, 2, *, Mengbin Ruan2, Peihong Wang3, Ming Peng2, *   

  1. 1Hainan University, Haikou 570228, China
    2Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China
    3Beijing Institute of Science and Technology Information, Beijing 100048, China
  • Received:2014-11-15 Accepted:2015-01-27 Online:2015-05-01 Published:2015-04-08
  • Contact: Sun Haiyan,Peng Ming
  • About author:

    ? These authors contributed equally to this paper

Abstract: The full-length thermostable α-amylase gene from a Bacillus licheniformis strain was cloned in a prokaryotic expression vector and a plant expression vector with GFP as reporter gene. Thermostable α-amylase was overexpressed in Escherichia coli by IPTG induction for 6 h at 28°C, protein bands at 55 kDa were detected with SDS-PAGE, and activity of the enzyme was confirmed by amylolysis experiments. The plant expression vector was transformed into tobacco via Agrobacterium tumefaciens mediated transformation. GFP expression was observed both in cytoplasm and vacuoles of the lower epidermis of tobacco by fluorescence microscopy. Staining of the tobacco leaf by I2-KI revealed that the thermostable α-amylase expressed in tobacco leaf had enzyme activity. Finally, the gene was introduced into Arabidopsis by the floral dip method and homozygous transgenic plants expressing α-amylase gene were identified. The results of this research provide further information for overexpression of thermostable α-amylase in transgenic plants.