Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Molecular Cloning of Differentially Expressed Genes in First Trap Leaf of Dionaea muscipula by Fluorescent Differential Display

형광 Differential Display법에 의한 파리지옥풀 포충잎트랩 특이발현 유전자 탐색

  • Kang, Kwon-Kyoo (Department of Horticulture, Hankyong National University) ;
  • Lee, Keun-Hyang (Department of Horticulture, Hankyong National University) ;
  • Park, Jin-Heui (Division of Biotechnology and Genetic Engineering, College of Life and Environmental Science, Korea University) ;
  • Hong, Kyong-Ei (Department of Horticulture, Hankyong National University)
  • Published : 2003.12.01
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Abstract

Fluorescent differential display (FDD) is a method for identifying differentially expressed genes in eukaryotic cells. The mRNA FDD technology works by systematic amplification of the 3' terminal regions of mRNAs. This method involve the reverse transcription using anchored primers designed to bind 5'boundary of the poly A tails, followed by polymerase chain reaction (PCR) amplification with additional upstream primers of arbitrary sequences. The amplified cDNA subpopulations are separated by denaturing polyacrylamide electrophoresis. To identify the genes involved in the development of first trap leaf, we applied a FDD method using mRNAs from leaf base, first trap leaf and flower tissue, respectively. We screened several genes that expressed specifically in first trap leaf. Nucleotide sequence analysis of these genes revealed that these were protease inhibitor (PI), myo-inositol-1-phosphate synthase and lipocalin-type prostaglandin D synthase. Northern blot analysis showed that these genes were expressed specifically in first trap leaf (in vivo and in vitro). FDD could prove to be useful for simultaneous scanning of transcripts from multiple cDNA samples and faster selection of differentially expressed transcripts of interest.

파리지옥풀 first trap leaf에만 발현하는 유전자군을 탐색하기 위하여 기내배양 식물체와 포충능력을 가진 3년생 실생주을 이용하여 각각의 포충잎 (leaf base), 꽃조직 (flower tissue) 및 포충잎트랩 (first trap leaf)으로부터 분리한 RNA로 Fluorescent differential display (FDD)를 실시하였다. First trap leaf특이발현 유전자 15개를 screening하여 염기서열을 분석하였다. 분리된 DNA들은 protease inhibitor (Pl), myo-inositol-1-phosphate synthase 및 lipocalin-type prostaglandin D syn-thase 유전자들과 매우 유사하였다. 또한 Northern blot분석 결과, 이들 유전자들이 first trap leaf에 특이적으로 발현하고 있는 것을 확인하였다 FDD방법은 세포, 조직 및 기관에 특이적으로 발현하고 있는 유전자들을 선발하는데 매우 유용한 수단으로 사용될 수 있다.

Keywords

References

  1. Abu-Abied M, Holland D (1994) Two newly isolated genes from citrus exhibit a different pattern of diumal expression and light response. Plant Mol Biol 26: 165-173 https://doi.org/10.1007/BF00039529
  2. Audsley N,Weaver RJ, Edwards JP (2002) Degradation of Manduca sexta allatostatin and allatotropin by proteases associated with the foregut of Lacanobia oleracea larvae. Peptides 23: 2015-2023 https://doi.org/10.1016/S0196-9781(02)00189-4
  3. Chun JA, Jin UH, Lee JW, Yi VB, Hyung NI, Kang MH, Pyee JH, Suh MC, Kang CW, Seo HY, Lee SW, Chung CH (2003) Isolation and characterization of a myo-inositol 1-phosphate synthase cDNA fromdeveloping sesame (Sesamum indicum L.) seeds: functional and differential expression, and salt-induced transcription during germination. Planta 216: 874-80
  4. de Obeso M, Caparros-Ruiz D, Vignols F, Puigdomenech P, Rigau J (2003) Characterisation of maize peroxidases having differential patterns of mRNA accumulation in relation to lignifying tissues. Gene 309: 23-33 https://doi.org/10.1016/S0378-1119(03)00462-1
  5. Facchini PJ, De Luca V (1995) Phloem-specific expression of tyro-sine/dopa decarboxylase genes and the biosynthesis of isoquino-line alkaloids in Opium poppy. Plant Cell 7: 1811-1821 https://doi.org/10.1105/tpc.7.11.1811
  6. George J (1962) Plants that eat insects. Readers Digest 2: 221-226
  7. Gonzalez RG, Haxo RS, Schleich T (1980) Mechanism of action of polymeric aurintricaboxylic acid, a potent inhibitor of protein-nucleic acid interactions. Biochemistry 19: 4299-4303 https://doi.org/10.1021/bi00559a023
  8. Gotoh K, Oishi M (2003) Screening of gene-associated polymor-phisms by use of in-gel competitive reassociation and EST (cDNA) array hybridization. Genome Res 13: 492-495 https://doi.org/10.1101/gr.434103
  9. Halitschke R, Gase K, Hui D, Schmidt DD, Baldwin IT (2003) Molecular interactions between the specialist herbivore manduca sexta (Lepidoptera, Sphingidae) and its natural host Nicotiana attenuata. VI. Microarray analysis reveals that most herbivore-specific transcriptional changes are mediated by fatty acid-amino acid conjugates. Plant Physiol 131: 1894-1902 https://doi.org/10.1104/pp.102.018184
  10. Hodick D, Sievers A (1989) On the mechanism of trap closure of Venus flytrap (Dionaea muscipula Ellis) Planta 179: 32-42 https://doi.org/10.1007/BF00395768
  11. lida R, Yasuda T, Tsubota E, Takatsuka H, Masuyama M, Matsuki T, Kishi K (2002) Five age-dependently expressed genes in mouse brain revealed by the fluorescence differential display-PCR technique. Exp Gerontol 37: 1121-1126 https://doi.org/10.1016/S0531-5565(02)00066-9
  12. Ito T, Kito K, Adati N, Mitsui Y, Hagiwara H, Sakaki Y (1994) Fluorescent differential display: arbitrarily primed RT-PCR finger-printing on an automated DNA sequencer. FEBS Lett 351: 231-236 https://doi.org/10.1016/0014-5793(94)00867-1
  13. Ito T, Sakaki Y (1996) Fluorescent differential display method for high speed scaning of tissue- or cell-specific transcripts. In methods In meolecular genetics, human molecular genetics edited by Adolph KW pp. 229-245. Academic Press, New York
  14. Kao YY, Harding SA, Tsai CJ (2002) Differential expression of two distinct phenylalanine ammonia-lyase genes in condensed tannin-accumulating and lignifying cells of quaking aspen. Plant Physiol 130: 796-807 https://doi.org/10.1104/pp.006262
  15. Liang R, Chan V, Chan TK, Wong T, Todd D (1992) Detection of immunoglobulin gene rearrangement in B-cell lymphomas by polymerasechain reactiongene amplification. Hematol Oncol 10: 149-154 https://doi.org/10.1002/hon.2900100304
  16. Louie M, Kondor N, DeWitt JG (2003) Gene expression in cadmium-tolerant Datura innoxia: detection and characterization of cDNAs induced in response to Cd^{2+}. Plant Mol Biol 52: 81-89 https://doi.org/10.1023/A:1023926225931
  17. Micchelli CA, The I, Selva E, Magila V, Perrimon N (2002) Rasp, a putative transmembrane acyltransferase, is required for hedgehog signaling. Development 129: 843-851
  18. Rachinsky A, Feldlaufer MF (2000) Responsiveness of honey bee (Apis meLlifera L.) corpora allata to allatoregulatory peptides from four insect species. J Insect Physiol 46: 41-46 https://doi.org/10.1016/S0022-1910(99)00100-6
  19. Roland J, Roland GN, Suzanne MC, John WB (1998) Identification and cloning of differenially expressed genes by long-distance differential display. Anal Biochem 259: 235-244 https://doi.org/10.1006/abio.1998.2653
  20. Salisbury FB, Ross CW (1985) Plant Physiology. Wadsworth Publ. Co. Belmony, CA. p 540
  21. Tamasloukht M, Sejalon-Delmas N, Kluever A, Jauneau A, Roux C, Becard G, Franken P (2003) Root factors induce mitochondrial-related gene expression and fungal respiration during the developmental switch from asymbiosis to presymbiosis in the arbuscular mycorrhizal fungus Gigaspora rosea. Plant Physiol 131: 1468-1478 https://doi.org/10.1104/pp.012898
  22. Torres GA, Lelandais-Briere C, Besin E, Jubier MF, Roche O, Mazubert C, Corre-Menguy F, Hartmann C (2003) Characterization of the expression of Phaseolus vulgaris OCT1, a dehydration-regulated gene that encodes a new type of phloem transporter. Plant Mol Biol 51: 341-349 https://doi.org/10.1023/A:1022014229899
  23. Wang J, Chen S (2002) Screening and identification of gastric adenocarcinoma metastasis-related genes using cDNA microar-ray coupledto FDD-PCR. J Cancer Res Clin Oncol 128: 547-553 https://doi.org/10.1007/s00432-002-0379-5
  24. Zhao GR, Liu JY, Du XM (2001) Molecular cloning and characterization of cotton cDNAs expressed in developing fiber cells. Biosci Biotechnol Biochem 65: 2789-2793 https://doi.org/10.1271/bbb.65.2789