Molecular Characterization of the Soybean L-Asparaginase Gene Induced by Low Temperature Stress

  • Cho, Chang-Woo (Department of Genetic Engineering, Dong-A University) ;
  • Lee, Hye-Jeong (Department of Genetic Engineering, Dong-A University) ;
  • Chung, Eunsook (Department of Genetic Engineering, Dong-A University) ;
  • Kim, Kyoung Mi (Department of Genetic Engineering, Dong-A University) ;
  • Heo, Jee Eun (Department of Genetic Engineering, Dong-A University) ;
  • Kim, Jung-In (School of Food and Life Science, Biohealth Products Research Center, Inje University) ;
  • Chung, Jongil (Department of Agronomy, Gyeongsang National University) ;
  • Ma, Youzhi (Institutes of Crop Breeding and Cultivation, China Academy of Agriculture Sciences) ;
  • Fukui, Kiichi (Department of Biotechnology, Graduate School of Engineering, Osaka University) ;
  • Lee, Dae-Won (Department of life Science, Dongguk University) ;
  • Kim, Doh-Hoon (Department of Genetic Engineering, Dong-A University) ;
  • Chung, Young-Soo (Department of Genetic Engineering, Dong-A University) ;
  • Lee, Jai-Heon (Department of Genetic Engineering, Dong-A University)
  • Received : 2006.11.30
  • Accepted : 2007.02.20
  • Published : 2007.06.30

Abstract

L-asparaginase (EC 3.5.1.1) catalyzes the hydrolysis of the amide group of L-asparagine, releasing aspartate and $NH_4{^+}$. We isolated a low temperature-inducible cDNA sequence encoding L-asparaginase from soybean leaves. The full-length L-asparaginase cDNA, designated GmASP1, contains an open reading frame of 1,258 bp coding for a protein of 326 amino acids. Genomic DNA blotting and fluorescence in situ hybridization showed that the soybean genome has two copies of GmASP1. GmASP1 mRNA was induced by low temperature, ABA and NaCl, but not by heat shock or drought stress. E. coli cells expressing recombinant GmASP1 had 3-fold increased L-asparaginase activity. A possible function of L-asparaginase in the early response to low temperature stress is discussed.

Keywords

Fluorescence In Situ Hybridization;Gene Expression;Low Temperature;Soybean;L-Asparaginase;Stress

Acknowledgement

Supported by : Dong-A University in Korea

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