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Journal of Plant Biology

, 49:364 | Cite as

Identification and characterization of highly expressed genes in suspension-cultured cells of sweet potato

  • Young Hwa Kim
  • Cheol Goo Hur
  • Yun Hee Shin
  • Jung Myung Bae
  • Young Sun Song
  • Gyung Hye HuhEmail author
Article

Abstract

Cultured plant cells have high potential in providing efficient and low-cost molecular farming systems for the large-scale production of commercially valuable recombinant proteins. As an initial aim at establishing an efficient expression system for suspension-cultured cells of food crops, we first obtained 1411 expressed sequence tags from a sweet potato cDNA library of exponential phase cells, and assembled them into 156 contigs and 1039 singletons. Five ESTs were selected as the most significantly abundant genes. These were transcripts for a cell wall development protein (cinnamyl alcohol dehydrogenase), a carbohydrate metabolite protein (glyceraldehyde-3-phosphate dehydrogenase), and a cell cycle regulator protein (small GTP binding protein Ran), as well as inorganic pyrophosphatase and cyclophilin. Comparisons were then made with the root and leaf EST libraries of sweet potato. Northern blot and RT-PCR analyses revealed that these five genes were strongly expressed in the suspension cells but not in the roots and leaves, thereby supporting the data obtained from the comparative analysis. This is the first reported comparison of the various EST libraries isolated from different cell types of the same plant species. These genes can now contribute to an applicable promoter for devising an efficient expression system from suspension-cultured cells.

Keywords

cinnamyl alcohol dehydrogenase expressed sequence tags glyceraldehyde-3-phosphate dehydrogenase small CTP binding protein Ran suspension-cultured cells sweet potato 

Abbreviations

CAD

cinnamyl alcohol dehydrogenase

DAS

days after subculture

dbEST

database expressed sequence tag

EST

expressed sequence tag

CAPDH

glyceraldehyde-3-phosphate dehydrogenase

MIPS

Munich Information Center for Protein Sequences

NR protein

non-redundant protein

Literature cited

  1. Birmbaum K, Shasha DE, Wang JY, Jung JW, Lambert GM, Galbraith DW, Benfey PN (2003) A gene expression map of theArabidopsis root. Science302: 1956–1960CrossRefGoogle Scholar
  2. Burke J, Davidson D, Hide W (1999) d2_cluster: A validated method for clustering EST and full length cDNA. Genome Res9: 1135–1142CrossRefPubMedGoogle Scholar
  3. Christoffels A, Gelder A, Greyling G, Miller R, Hide T, Hide W (2001) STACK: Sequence tag alignment and consensus knowl-edgebase. Nucl Acids Res29: 234–238CrossRefPubMedGoogle Scholar
  4. Crookshanks M, Emmersen J, Welinder KG, Nielsen KL (2001) The potato tuber transcriptome: Analysis of 6077 expressed sequence tags. FEBS Lett506: 123–126CrossRefPubMedGoogle Scholar
  5. Doran PM (2000) Foreign protein production in plant tissue cultures. Curr Opin Biotechnol11: 199–204CrossRefPubMedGoogle Scholar
  6. Ewing B, Millier L, Wendl MC, Green P (1998) Base-calling of automated sequencer traces using Pred. I. Accuracy assessment. Genome Res8: 175–185PubMedGoogle Scholar
  7. Fischer R, Stoger E, Schillberg S, Christou P, Twyman RM (2004) Plant-based production of biopharmaceuticals. Curr Opin Plant Biol7: 152–158CrossRefPubMedGoogle Scholar
  8. Hellwing S, Drossard J, Twyman RM, Fischer R (2004) Plant cell cultures for the production of recombinant proteins. Nat Biotechnol22: 1415–1422CrossRefGoogle Scholar
  9. Hide W, Burke J, Davidson D (1994) Biological evaluation of d2, an algorithm for high performance sequence comparison. J Comput Biol1: 199–215CrossRefPubMedGoogle Scholar
  10. Kim DJ (2004) A study of cotyledon senescence in cucumber (Cucumis sativus L.) based on expressed sequence tags and gene expression. J Plant Biol47: 244–253CrossRefGoogle Scholar
  11. Kim SK, Kwak SS, Jung KH, Min SR, Park IH, Liu JR (1994) Selection of plant cell lines for high yields of peroxidase. Kor Biochem J27: 132–137Google Scholar
  12. Kim YH, Cho EH, Kwak SS, Kwon SK, Bae JM, Lee BR, Meen Bl, Huh GH (2004) Alterations in intracellular and extracellular activities of antioxidant enzymes during suspension culture of sweet potato. Phytochem65: 2471–2476CrossRefGoogle Scholar
  13. Kwon TH, Kim YS, Lee JH, Yang MS (2003) Production and secretion of biologically active human granulocyte-macrophage colony stimulating factor in transgenic tomato suspension cultures. Biotechnol Lett25: 1571–1574CrossRefPubMedGoogle Scholar
  14. Li HY, Cao K, Zheng Y (2003) Ran in the spindle checkpoint: A new function for a versatile CTPase. Trends Cell Biol13: 553–557CrossRefPubMedGoogle Scholar
  15. Matsuoka K, Demura T, Galis I, Horiguchi T, Sasaki M, Tashiro G, Fukuda H (2004) A comprehensive gene expression analysis toward the understanding of growth and differentiation of tobacco BY-2 cells. Plant Cell Physiol45: 1280–1289CrossRefPubMedGoogle Scholar
  16. Mewes HW, Frichman D, Guidener W, Mannhaupt G, Mayer K, Mokrejs M, Morgenstern B, Munsterkotter M, Rudd S, Weil B (2002) MIPS: A database for genomes and protein sequences. Nucl Acids Res30: 31–34CrossRefPubMedGoogle Scholar
  17. Miller R, Christoffels A, Gopalakrishnan C, Burke J, Ptitsyn AA, Broveak TR, Hide W (1999) A comprehensive approach to clustering of expressed human gene sequence: The sequence tag alignment and consensus knowledgebase. Genome Res9: 1143–1155CrossRefPubMedGoogle Scholar
  18. Mitchell HJ, Hall JL, Barber MA (1994) Elicitor-induced cinnamyl alcohol dehydrogenase activity in lignifying wheat (Triticum aestivum L.) leaves. Plant Physiol104: 551–556PubMedGoogle Scholar
  19. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assay with tobacco tissue cultures. Physiol Plant15: 473–497CrossRefGoogle Scholar
  20. Robert A, Gruissem W (1994) A small nuclear GTP-binding protein from tomato suppresses aSchizosaccharomyces pombe cell-cycle mutant. Proc Natl Acad Sci USA91: 5863–5867CrossRefGoogle Scholar
  21. Sauter M, Kende H (1992) Levels of a-glucan and lignin in elongating internodes of deepwater rice. Plant Cell Physiol33: 1089–1097Google Scholar
  22. Shin YJ, Hong SY, Kwon TH, Jang YS, Yang MS (2003) High level of expression of recombinant human granulocyte-macrophage colony stimulating factor in transgenic rice cell suspension culture. Biotechnol Bioengr82: 778–783CrossRefGoogle Scholar
  23. Smith ML, Mason HS, Shuler ML (2002) Hepatitis B surface antigen (HBsAg) expression in plant cell culture: Kinetics of antigen accumulation in batch culture and its intracellular form. Biotechnol Bioengr80: 812–822CrossRefGoogle Scholar
  24. Spang R, Vingron M (2000) Limits of homology detection by pair-wise sequence comparison. Bioinformatics17: 338–342CrossRefGoogle Scholar
  25. Suh MC, Kim MJ, Hur CG, BaeJM, Park Yl, Chung CH, Kang CW, Ohlrogge JB (2003) Comparative analysis of expressed sequence tags fromSesamum indicum andArabidopsis thaliana developing seeds. Plant Mol Biol52: 1107–1123CrossRefPubMedGoogle Scholar
  26. Torres E, Vaquera C, Nicholson L, Sack M, Stoger E, Drossard J, Christou P, Fischer R, Perrin Y (1999) Rice cell culture as an alternative production system for functional diagnostic and therapeutic antibodies. Transgen Res8: 441–449CrossRefGoogle Scholar
  27. Twyman RM, Stoger E, Schillberg S, Christou P, Fischer R (2003) Molecular farming in plants: Host systems and expression technology. Trends Biotechnol21: 570–578CrossRefPubMedGoogle Scholar
  28. You MK, Hur CG, Ahn YS, Suh MC, Jeong BC, Shin JS, Bae JM (2003) Identification of genes possibly related to storage root induction in sweet potato. FEBS Lett536: 101–105CrossRefPubMedGoogle Scholar

Copyright information

© The Botanical Society of Korea 2006

Authors and Affiliations

  • Young Hwa Kim
    • 1
    • 2
  • Cheol Goo Hur
    • 3
  • Yun Hee Shin
    • 3
  • Jung Myung Bae
    • 4
  • Young Sun Song
    • 2
  • Gyung Hye Huh
    • 1
    Email author
  1. 1.College of General EducationInje UniversityGimhaeKorea
  2. 2.School of Food and Life Science, BPRCInje UniversityGimhaeKorea
  3. 3.Plant Genome Laboratory, Genome Research CenterKRIBBDaejeonKorea
  4. 4.School of Life Sciences and BiotechnologyKorea UniversitySeoulKorea

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