Cloning and Nucleotide Sequencing of a Partial Glutamate Decarboxylase Gene from Arabidopsis thaliana cDNA Library

애기장대 cDNA library로부터 Glutamate Decarboxylase 유전자의 부분 클로닝 및 서열분석

  • Published : 2001.02.01

Abstract

In order to study the molecular mechanism of $\gamma$-aminobutyric acid (GABA) production in plants, we cloned and sequenced a partial glutamate decarboxylase (GAD) cDNA from the Arabidopsis thaliana cDNA library, using primers targeted at highly conserved sequences of the petunia GAD gene. The cDNA fragment was inserted into TA cloning vector with T7 promoter and the recombinant plasmid obtained was used to transform E. coli. The plasmid DNA purified from the transformed E. coli was digested with EcoRI and the presence of the insert was confirmed. Nucleotide sequence analysis showed that the fragment is a partial Arabidopsis thaliana GAD gene and that the sequence showed 98% and 78% identity to the region of the putative Arabidopsis thaliana GAD sequences deposited in GenBank, Accession nos: U46665 and U10034, respectively. The amino acid sequence deduced from the partial Arabidopsis thaliana GAD gene showed 99% and 91% identities to the GAD sequences deduced from the genes of the U46665 and U10034, respectively. The partial cDNA sequence determined may facilitate the study of the molecular mechanism of GABA metabolism in plants.

References

  1. Phytochem. v.29 Metabolism enzymology and possible roles of 4-aminobutyrate in higher plants Satyanarayan, V.;P. M. Nair
  2. Plant Physiol. v.115 The metabolism and functions of γ-aminobutyric acid Bown, A. W.;B. J. Shelp
  3. Trends in Plant Sci. v.3 Calmodulin, calmodulin-related proteins and plant responses to the environment Snedden, W. A.;H. Fromm
  4. Plant Physiol. v.108 Molecular and biochemical analysis of calmodulin : Interactions with the calmodulin-binding domain of plant glutamate decarboxylase Arazi, T.;G. Baum;W. A. Snedden;B. J. Shelp;H. Fromm
  5. EMBO J. v.15 Calmodulin binding to glutamate decarboxylase is required for regulation of glutamate and GABA metabolism and normal development in plants Baum. G.;S. Lev-Yadun;Y. Fridmann;T. Arazi;H. Katsnelson;M. Zik;H. Fromm
  6. Trends in Plant Sci. v.4 Metabolism and functions of gamma-aminobutyric acid Shelp, B. J.;A. W. Bown;M. D. McLean
  7. Proc. Natl. Acad. Sci. USA v.89 Wind-induced plant motion immediately increases cytosolic calcium Knight, M. R.;S. M. Smith;A. J. Trewavas
  8. Plant Physiol. v.90 Salt stress-induced cytoplasmic acidification and vacuolar alkalization in Nitellopsis obtusa cells Katsuhara, M.;K. Kuchitsu;K. Takeshige;M. Tazawa
  9. Plant Physiol. v.108 Calcium/calmodulin activation of soybean glutamate decarboxylase Snedden, W. A.;T. Arazi;H. Fromm;B. J. Shelp
  10. Plant Cell v.6 Analysis of a soluble calmodulin binding protein from fava bean roots: Identification of glutamate decarboxylase as a calmodulin-activated enzyme Ling, V.;W. A. Snedden;B. J. Shelp;S. M. Assmann
  11. J. Biol. Chem. v.271 Activation of a recombinant petunia glutamate decarboxylase by calcium/calmodulin or by a monoclonal antibody which recognizes the calmodulin binding domain Snedden, W. A.;N. Koutsia;G. Baum;H. Fromm
  12. Plant Mol. Biol. v.37 Two isoforms of glutamate decarboxylase in Arabidopsis are regulated by calcium/calmodulin and differ in organ distribution Zik, M.;T. Arazi;W. A. Snedden;H. Fromm
  13. Cell v.60 Rain-, wind-, and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis Braam, J.;R. W. Davis
  14. J. Biol. Chem. v.270 Identification of a novel divergent calmodulin isoform from soybean which has differential ability to activate calmodulin-dependent enzymes Lee, S. H.;J. C. Kim;M. S. Lee;W. D. Heo;H. Y. Seo;H. W. Yoon;J. C. Hong;S. Y. Lee;J. D. Bahk;I. Hwang;M. J. Cho
  15. Receptor Pharmacology and Function GABA receptors Krogsgaard-Larsen, P.;M. Williams(ed.);R. A. Glennon(ed.);P.M.W.M. Timmermans(ed.)
  16. J. Biol. Chem. v.270 Brain L-glutamate decarboxylase Bao, J.;W. Y. Cheung;J. Y. Wu
  17. Am. J. Psychiatry v.155 Increased glutamatergic neurot ransmission and oxidative stress after alcohol withdrawal Tsai, G. E.;P. Ragan;R. Chang;S. Chen;V. H. Linnoila;J. T. Coyle
  18. Kor. J. Biotechnol. Bioeng. v.15 Production of the quality germinated brown rices containing high γ-aminobutyric acid by chitosan application Oh, S.-H.;W.-G. Choi
  19. Proc. Natl. Acad. Sci. USA v.74 DNA sequencing with chain-terminating inhibitors Sanger, F.;S. Nicklen;A. R. Coulson
  20. Korean J. Food. Sci. Technol. v.24 Changes in γ-aminobutyric acid(GABA) and the main constituents by treatment conditions and of anaerobically treated green tea leaves Chang, J. S.;B. S. Lee;Y. G. Kim
  21. Korean Soci. Crop Sci. v.43 Effect of anaerobic treatment on carbohydrate-hydrolytic enzyme activities and free amino acid contents in barly malt Yun, S. J.;K. G. Choi;J. K. Kim
  22. J. Korean Soc. Food Sci. Nutr. v.29 Investigation of γ-aminobutyric acid in Chinese cabbages and effects of the cabbage diets on lipid metabolism and liver function of rats administered with ethanol Cha, Y.-S.;S.-H. Oh