Science in China Series C: Life Sciences

, Volume 42, Issue 3, pp 274–280 | Cite as

Expression ofArabidopsis tryptophan biosynthetic pathway genes: effect of the 5′ coding region of phosphoribosylanthranilate isomerase gene

  • Yikun He
  • Xinfang Liu
  • Jiayang Li


There are three non-allelic isogenes encoding phosphoribosylanthranilate isomerase (PAI) inArabidopsis thaliana. The expression plasmids were constructed by fusion of the GUS reporter gene to the three PAI promoters with or without the 5′ region encoding PAI N-terminal polypeptides and transferred into Arabidopsis plants byAgrobacterium tumefaciens. Analysis of GUS activity revealed that the PAI 5′ coding region was necessary for high expression of GUS activity. GUS activity in transgenic plants transformed with the expression plasmids containing the 5′ coding region of PAH or PAI3 was 60–100-fold higher than that without the corresponding 5′ region. However, the effect of 5’ coding region of PAI2 gene on the GUS activity was very small (only about 1 time difference). The GUS histochemical staining showed a similar result as revealed by GUS activity assay. It was expressed in the mesophyll cells and guard cells, but not in the epidermic cells, indicating that the N-terminal polypeptides encoded by the 5′ region of PAI genes have the function of PTP.


phosphoribosylanthranilate isomerase(PAI) tryptophan biosynthetic pathway gene expression and regulation plastid targeting peptide (PTP) Arabidopsis thaliana 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Highfield, P. E., Ellis, R. J., Synthesis and transport of the small subunit of chloroplast ribulose bisphosphate carboxylase,Nature, 1978, 271(2): 420.CrossRefGoogle Scholar
  2. 2.
    Keegstra, K., Olsen, L. J., Chloroplastic precursors and their transport across the envelope membranes,Annu. Rev. Plant Physiol. Plant Mol. Biol., 1989, 40: 471.CrossRefGoogle Scholar
  3. 3.
    Roberts, B. E., Paterson, B. M., Efficient translation of tobacco mosaic virus RNA and rabbit globulin 9 s RNA in a cell-free system from commercial wheat germ,Proc. Natl. Acad. Sci. USA, 1973, 70: 2330.CrossRefPubMedGoogle Scholar
  4. 4.
    Ellis, R. J., Protein synthesis by isolated chloroplasts,J. Biochemi. Biophs. Acta, 1977, 463: 185.Google Scholar
  5. 5.
    Chua, N. H., Schmidt, G. W., Transport of proteins into mitochondria and chloroplasts,J. Cell. Biol., 1979, 81: 461.CrossRefPubMedGoogle Scholar
  6. 6.
    Li, J., Zhao, J., Rose, A. B. et al., Arabidopsis phosphoribosylanthranilate isomerase: Molecular genetic analysis of triplicate tryptophan pathway genes,Plant Cell, 1995, 7: 447.CrossRefPubMedGoogle Scholar
  7. 7.
    Zhao, J., Last, R. L., Immunological characterization and chloroplast localization of the tryptophan biosynthetic enzymes of the flowering plant Arabidopsis thaliana,J. Biol. Chem., 1995, 270: 6081.CrossRefPubMedGoogle Scholar
  8. 8.
    Pruitt, K. P., Last, R. L., Expression pattern duplicate tryptophan synthase gene in Arabidopsis thaliana,Plant Physiol., 1993, 102: 1019.CrossRefPubMedGoogle Scholar
  9. 9.
    Jefferson, R. A., Assaying chemeric genes in plants: The Gus gene fusion system,Plant Mol. Biol. Rep., 1987, 5: 387.CrossRefGoogle Scholar
  10. 10.
    Bradford, M. M., A rapid and sensitive method for the quantitation of microgram quantites of protein utilizing the principle of protein-dye binding,Anal. Biochem., 1976, 72: 248.CrossRefPubMedGoogle Scholar
  11. 11.
    Stomp, A. M., Histochemical localization of β-Glucuronidase, inGUS protocols: Using the GUS Gene as a Reporter of Gene Expression (ed. Gallagher, S. R.), San Diego: Academic Press, Inc., 1992, 103–114.Google Scholar
  12. 12.
    Rose, A. B., Casselman, A. L., Last, R. L., A phosphoribosyl anthranilate transferase gene is defective in blue fluorescent Arabidopsis thaliana tryptophan mutants,Plant Physiol., 1992, 100: 582.CrossRefPubMedGoogle Scholar
  13. 13.
    Radwanski, E. R., Last, R. L., Tryptophan biosynthesis and metabolism: Biochemical and molecular genetics,Plant Cell, 1995, 7: 921.CrossRefPubMedGoogle Scholar
  14. 14.
    Odell, J. T., Nagy, F., Chua, N. H., Identification of DNA sequences required for activity of the cauliflower mosaic virus 35 S promoter,Nature, 1985, 313: 810.CrossRefPubMedGoogle Scholar
  15. 15.
    Rose, A.B., Last, R. L., Introns act post-transcriptionally to increase expression of theArabidopsis thaliana tryptophan pathway gene PAT1,The Plant Journal, 1997, 11(3); 455.CrossRefPubMedGoogle Scholar

Copyright information

© Science in China Press 1999

Authors and Affiliations

  • Yikun He
    • 1
    • 2
  • Xinfang Liu
    • 1
    • 2
  • Jiayang Li
    • 1
    • 2
  1. 1.Plant Biotechnology Laboratory, Institute of GeneticsChinese Academy of SciencesChina
  2. 2.Chinese CenterDe Montfort University, Norman Borlaug Institute for Plant Science ReseachBeijingChina

Personalised recommendations