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PHD : Parasites, Hosts and Diseases

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Original Article

Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii

The Korean Journal of Parasitology 2011;49(4):341-347.
Published online: December 16, 2011

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Corresponding author (hhkong@mail.knu.ac.kr)
• Received: July 28, 2011   • Revised: September 6, 2011   • Accepted: September 9, 2011

© 2011, Korean Society for Parasitology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii
Korean J Parasitol. 2011;49(4):341-347.   Published online December 16, 2011
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Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii
Korean J Parasitol. 2011;49(4):341-347.   Published online December 16, 2011
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Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii
Image
Fig. 1 Comparison of gene expression by microarray and KOG analysis. Cyst up-regulated genes showed higher percentages in A, D, T, W, U, O, and F articles. In J, C, and Z (cytoskeleton) articles, cyst genes were down-regulated.
Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii
KOG category No. of genes (%)
Cyst-high Troph-high Information storage and processing  J Translation, ribosomal structure and biogenesis 28 (3.99) 135 (15.72)  A RNA processing and modification 27 (3.85) 22 (2.56)  K Transcription 10 (1.43) 11 (1.28)  L Replication, recombination and repair 1 (0.14) 4 (0.47)  B Chromatin structure and dynamics 3 (0.43) 9 (1.05) Metabolism  C Energy production and conversion 14 (2.00) 67 (7.80)  G Carbohydrate transport and metabolism 12 (1.71) 26 (3.03)  E Amino acid transport and metabolism 9 (1.28) 25 (2.91)  F Nucleotide transport and metabolism 17 (1.40) 13 (1.51)  H Coenzyme transport and metabolism 0 (0.00) 11 (1.28)  I Lipid transport and metabolism 12 (1.71) 23 (2.68)  P Inorganic ion transport and metabolism 7 (1.00) 5 (0.28)  Q Secondary metabolites biosynthesis, transport and metabolism 5 (0.71) 12 (1.40) Cellular process and signaling  D Cell cycle control, cell division, chromosome partitioning 7 (1.00) 5 (0.58)  Y Nuclear structure 0 (0.00) 1 (0.12)  V Defense mechanisms 3 (0.43) 2 (0.23)  T Signal transduction mechanisms 36 (5.14) 23 (2.68)  M Cell wall/membrane/envelope biogenesis 3 (0.43) 0 (0.00)  N Cell motility 0 (0.00) 0 (0.00)  Z Cytoskeleton 6 (0.86) 39 (4.54)  W Extracellular structures 16 (2.28) 12 (1.40)  U Intracellular trafficking, secretion, and vesicular transport 25 (3.57) 15 (1.75)  O Posttranslational modification, protein turnover, chaperones 39 (5.56) 30 (3.49) Poorly characterized  R General function prediction only 33 (4.71) 37 (4.31)  S Function unknown 7 (1.00) 13 (1.51) Not classified 381 (54.35) 319 (37.14) Total 701 (100.00) 859 (100.00) ID Probe comment Fold difference
Primer sequence Microarray Real-time PCR NC0115-T3 Cysteine proteinase; ACCP2 282.38 1,384 F: 5´-AAC AGC ACG CTC GTT TCC CTC T-3´ R: 5´-GTA GTT GGC CTC CGT CAT GAG CTT-3´ ACL00008706 Catalase related cluster 56.30 11.0 F: 5´-ATC GCA CAC ACA CCA CAC TTT C-3´ R: 5´-CAA CGG AGC ACA AGA GAA CAA A-3´ NC0392-T3 Proteophosphoglycan ppg4 31.21 137.19 F: 5´-TCA TCA TCA CCT CGC TAC ACC A-3´ R: 5´-TAG TTG CAG GTG CGT TCG AGA-3´ NC0903-T3 Arginase II 9.27 20.25 F: 5´-CTC AAG GAG CTA GAC GGT GGA A-3´ R: 5´-TAG GTG TCG AGA AGG CCA GAG A-3´ AC-T0782-T3 Cyst specific protein CSP21 5.19 24.95 F: 5´-GAG AAG TAC GGC AAC GTG GAG A-3´ R: 5´-GCA TAG AGC AGG CGT TCA ACA-3´ ACL00008536 Dipeptidyl peptidase IV cluster 4.64 4.06 F: 5´-ACT GGC GAC ACA CCG TCT ATC T-3´ R: 5´-TGC GCG AGG AAT CAA CAA A-3´ NC0224 Protein serine/threonine kinase 3.85 8.17 F: 5´-CCA CAT CAA CCA AAT CAC CGA-3´ R: 5´-CGC ATA ACC GAA CAT CAT CCA-3´ ACL00001979 Ubiquitin 3.69 13.27 F: 5´-ACA CAC AAG CCA GCA ATG CA-3´ R: 5´-AAG TGG TGG AGG AAA CAC GCT-3´ AC-C0063 Heat shock protein 2.8 3.41 F: 5´-GAC CGA CGG CAA TAA ACT CGT-3´ R: 5´-TCG CTG CAC CAA CAG ATT CTC-3´ ID Probe comment KOG Fold (Cyst/troph) NC0115-T3 Cysteine proteinase; ACCP2 O 282.38 NC0961-T3 Cysteine proteinase; ACCP2 O 273.54 AC-T0581-T3 Similar to elaC homology R 80.82 ACE00000784 - - 73.39 ACL00003940 Herpes_BLLF1 multi-domain protein - 60.94 ACL00008706 Catalase related cluster AR 56.30 NC0692-T3 Extracellular endoglucanase WV 54.94 NC0680-T3 - - 53.32 ACL00002157 - - 48.23 NC0642-T3 - - 41.34 NC0757-T3 - - 38.91 ACL00003760 - - 36.86 ACL00009010 - - 36.09 ACL00004491 - - 33.26 NC0678-T3 - - 31.73 NC0457-T3 - - 31.72 NC0392-T3 Proteophosphoglycan ppg4 - 31.21 AC-C0617-T3 SJCHGC01975 protein O 29.79 ACL00008637 Novel protein related cluster U 27.80 NC0271-T3 Salivary gland secretion 1 CG3047-PA - 27.04 NC0289-T3 - - 26.25 NC0423-T3 Proteophosphoglycan 5 - 23.32 ACL00004246 - - 23.31 NC0956-T3 Similar to serine/arginine repetitive matrix 1 - 22.54 NC0802-T3 Proteophosphoglycan 5 WV 21.72 ACL00002714 Dynamin B precursor related cluster UR 21.23 NC0276-T3 Similar to conserved hypothetical protein T 20.90 ACE00001346 Similar to molecular chaperone (HSP90 family) O 20.34 NC0283-T3 Proteophosphoglycan ppg4 - 19.68 ACL00008639 Herpes_gp2 multi-domain protein - 19.22 NC0994-T3 Proteophosphoglycan 5 - 19.13 NC0807-T3 Antifreeze glycoprotein - 18.43 NC0791-T3 Hypothetical protein Kpol_1072p54 DO 17.66 ACL00002613 - - 17.13 NC1140-T3 Proteophosphoglycan ppg4 - 16.94 NC0582-T3 - - 16.92 ACL00002232 - - 16.50 ACL00002108 Transcription factor TFIIF K 15.49 NC0747-T3 Pseudogene - 15.27 NC0325-T3 Proteophosphoglycan ppg4 - 15.07 NC1103-T3 Proteophosphoglycan ppg4 - 14.95 AC-T0529-T3 - - 14.77 ACL00002483 - - 14.58 AC-T0879-T3 Iron-sulfur cluster assembly fam... CU 14.54 NC0896-T3 Proteophosphoglycan ppg4 - 14.53 ACL00001119 trEST Trithorax protein related cluster R 13.83 NC0987-T3 Palmitoyl-protein thioesterase 3 IO 13.77 NC0591-T3 Similar to scavenger receptor class F, member 2, partial WV 13.70 NC0780-T3 Hemerythrin HHE cation binding domain-containing protein - 13.63 ACL00008833 Extensin_2 multi-domain protein T 12.98 ACL00008542 - - 12.47 ACL00009103 - - 12.34 ACL00009220 - - 11.76 NC0721-T3 LigA - 11.57 NC0626-T3 Proteophosphoglycan 5 A 11.57 ACL00000192 Ubiquitin OR 11.03 NC1049-T3 Antifreeze glycoprotein - 10.89 ACL00008353 Tymo_45kd_70kd multi-domain protein WV 10.73 NC0528-T3 BTB/POZ domain-containing protein - 10.62 ACL00002099 Putative 5-3 exoribonuclease related cluster LA 10.47 ACL00008536 Dipeptidyl peptidase IV related cluster O 10.46 ACL00003598 trEST Herpes_UL56 multi-domain protein - 10.35 ACL00004676 - - 10.27 AC-T0408-T3 Histone H3 B 10.16 NC0574-T3 Hypothetical protein MGG_00576 - 10.10 ACL00000775 Unknown (eubacterial similarity) - 10.05
Table 1. KOG analysis of cyst and trophozoite genes up-regulated more than 2 folds
Table 2. Real-time PCR analysis of selected genes based on the microarray results
Table 3. The list of genes that showed high expression changes more than 10 fold in cysts