Parasites, Hosts and Diseases

"encystation"

Original Articles

Identification of essential genes for Acanthamoeba castellanii excystation during encystation and excystation: Min-Jeong Kim, Hye-Jeong Jo, Fu-Shi Quan, Ki Back Chu, Hyun-Hee Kong, Eun-Kyung Moon; Parasites Hosts Dis 2024;62(4):399-407.
Published online November 22, 2024; DOI: https://doi.org/10.3347/PHD.24062

Acanthamoeba is an opportunistic pathogen that causes Acanthamoeba keratitis, granulomatous amoebic encephalitis, and other cutaneous diseases. The life cycle of Acanthamoeba consists of 2 stages of trophozoites and cysts. Under adverse environmental conditions, Acanthamoeba encysts, while the conditions become favorable for growth, it reverts to the trophozoite form. Acanthamoeba excystation is crucial for its proliferation and can lead to recurrent infections after incomplete treatment. To identify the factors involved in excystation, A. castellanii was subjected to either encystation- or excystation-inducing conditions, and gene expression profiles were compared using mRNA sequencing. A. castellanii samples were collected at 8 h intervals for analysis under both conditions. Differentially expressed gene analysis revealed that 1,214 and 1,163 genes were upregulated and downregulated, respectively, by more than 2-fold during early excystation. Five genes markedly upregulated in early excystation (ACA1_031140, ACA1_032330, ACA1_374400, ACA1_275740, and ACA1_112650) were selected, and their expression levels were confirmed via real-time PCR. Small interfering RNA (siRNA) targeting these 5 genes was transfected into Acanthamoeba and gene knockdown was validated through real-time PCR. The silencing of ACA1_031140, ACA1_032330, ACA1_374400, and ACA1_112650 inhibited excystation and suggested that these genes might be essential for excystation. Our findings provide valuable insights for suppressing Acanthamoeba proliferation and recurrence.

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Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher, Sébastien Pomel
Pathogens.2025; 14(3): 268. CrossRef
Integrating molecular pathogenesis and predictive modelling in granulomatous amoebic encephalitis due to Acanthamoeba : a digital twin framework for personalised therapy
Ruqaiyyah Siddiqui, David Lloyd, Naveed Ahmed Khan
Infectious Diseases.2025; : 1. CrossRef
Microarray-based characterization of airway inflammation induced by environmental Acanthamoeba exposure versus the ovalbumin-alum model
Shin Ae Kang, Hak Sun Yu
Parasites, Hosts and Diseases.2025; 63(4): 327. CrossRef

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Sirtinol Supresses Trophozoites Proliferation and Encystation of Acanthamoeba via Inhibition of Sirtuin Family Protein: So-Young Joo, Ja Moon Aung, Minsang Shin, Eun-Kyung Moon, Hyun-Hee Kong, Youn-Kyoung Goo, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2022;60(1):1-6.
Published online February 23, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.1.1

Abstract
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The encystation of Acanthamoeba leads to the development of metabolically inactive and dormant cysts from vegetative trophozoites under unfavorable conditions. These cysts are highly resistant to anti-Acanthamoeba drugs and biocides. Therefore, the inhibition of encystation would be more effective in treating Acanthamoeba infection. In our previous study, a sirtuin family protein—Acanthamoeba silent-information regulator 2-like protein (AcSir2)—was identified, and its expression was discovered to be critical for Acanthamoeba castellanii proliferation and encystation. In this study, to develop Acanthamoeba sirtuin inhibitors, we examine the effects of sirtinol, a sirtuin inhibitor, on trophozoite growth and encystation. Sirtinol inhibited A. castellanii trophozoites proliferation (IC50=61.24 μM). The encystation rate of cells treated with sirtinol significantly decreased to 39.8% (200 μM sirtinol) after 24 hr of incubation compared to controls. In AcSir2-overexpressing cells, the transcriptional level of cyst-specific cysteine protease (CSCP), an Acanthamoeba cysteine protease involved in the encysting process, was 11.6- and 88.6-fold higher at 48 and 72 hr after induction of encystation compared to control. However, sirtinol suppresses CSCP transcription, resulting that the undegraded organelles and large molecules remained in sirtinol-treated cells during encystation. These results indicated that sirtinol sufficiently inhibited trophozoite proliferation and encystation, and can be used to treat Acanthamoeba infections.

Citations

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Songlan Gao, Honglei Zhang, Na Li, Lijuan Zhang, Zhe Zhu, Changlu Xu
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Kalpana Pandya, Krishnashish Roul, Avanish Tripathi, Sateesh Belemkar, Anshuman Sinha, Meryem Erol, Devendra Kumar
ACS Chemical Neuroscience.2025; 16(12): 2178. CrossRef
Human Conjunctival Transcriptome in Acanthamoeba Keratitis: An Exploratory Study
Gerami D. Seitzman, Jeremy D. Keenan, Thomas M. Lietman, Kevin Ruder, Lina Zhong, Cindi Chen, YuHeng Liu, Danny Yu, Thomas Abraham, Armin Hinterwirth, Thuy Doan
Cornea.2024; 43(10): 1272. CrossRef
Comparative cytotoxicity of Acanthamoeba castellanii-derived conditioned medium on human corneal epithelial and stromal cells
Abdullah Alhazmi, Laura E. Sidney, Andy Hopkinson, Hany M. Elsheikha
Acta Tropica.2024; 257: 107288. CrossRef
Biological characteristics and pathogenicity of Acanthamoeba
Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
Frontiers in Microbiology.2023;[Epub] CrossRef

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Transcriptomic Features of Echinococcus granulosus Protoscolex during the Encystation Process: Junjie Fan, Hongye Wu, Kai Li, Xunuo Liu, Qingqing Tan, Wenqiao Cao, Bo Liang, Bin Ye; Korean J Parasitol 2020;58(3):287-299.
Published online June 26, 2020; DOI: https://doi.org/10.3347/kjp.2020.58.3.287

Cystic echinococcosis (CE) is a zoonotic infection caused by Echinococcus granulosus larvae. It seriously affects the development of animal husbandry and endangers human health. Due to a poor understanding of the cystic fluid formation pathway, there is currently a lack of innovative methods for the prevention and treatment of CE. In this study, the protoscoleces (PSCs) in the encystation process were analyzed by high-throughput RNA sequencing. A total of 32,401 transcripts and 14,903 cDNAs revealed numbers of new genes and transcripts, stage-specific genes, and differently expressed genes. Genes encoding proteins involved in signaling pathways, such as putative G-protein coupled receptor, tyrosine kinases, and serine/threonine protein kinase, were predominantly up-regulated during the encystation process. Antioxidant enzymes included cytochrome c oxidase, thioredoxin glutathione, and glutathione peroxidase were a high expression level. Intriguingly, KEGG enrichment suggested that differentially up-regulated genes involved in the vasopressin-regulated water reabsorption metabolic pathway may play important roles in the transport of proteins, carbohydrates, and other substances. These results provide valuable information on the mechanism of cystic fluid production during the encystation process, and provide a basis for further studies on the molecular mechanisms of growth and development of PSCs.

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First comparative proteomic and in vitro behavioral study of Echinococcus granulosus metacestodes in Felis catus
Andrea Maglioco, Vanesa V. Miana, María Pía Valacco, Facundo A. Agüero, María Laura Gertiser, Héctor G. Avila, Melisa S. Barbery Venturi, Oscar Jensen, Alejandra Y. Juárez Valdez, Elio A. Prieto González, Alicia G. Fuchs
Frontiers in Veterinary Science.2025;[Epub] CrossRef
Genome-wide transcriptome analysis of Echinococcus multilocularis larvae and germinative cell cultures reveals genes involved in parasite stem cell function
Michaela Herz, Magdalena Zarowiecki, Leonie Wessels, Katharina Pätzel, Ruth Herrmann, Christiane Braun, Nancy Holroyd, Thomas Huckvale, Monika Bergmann, Markus Spiliotis, Uriel Koziol, Matthew Berriman, Klaus Brehm
Frontiers in Cellular and Infection Microbiology.2024;[Epub] CrossRef
In vitro and in silico scolicidal effect of sanguinarine on the hydatid cyst protoscoleces
Elham Hassanzadeh, Shahram Khademvatan, Behzad Jafari, Abbas Jafari, Elham Yousefi, Sheikh Arslan Sehgal
PLOS ONE.2023; 18(10): e0290947. CrossRef
Transcriptome analysis of Echinococcus granulosus sensu stricto protoscoleces reveals differences in immune modulation gene expression between cysts found in cattle and sheep
Ismael Pereira, Christian Hidalgo, Caroll Stoore, María Soledad Baquedano, Carolina Cabezas, Macarena Bastías, Aníbal Riveros, Claudio Meneses, Martín Cancela, Henrique Bunselmeyer Ferreira, Leonardo Sáenz, Rodolfo Paredes
Veterinary Research.2022;[Epub] CrossRef
Chromosome-scale Echinococcus granulosus (genotype G1) genome reveals the Eg95 gene family and conservation of the EG95-vaccine molecule
Pasi K. Korhonen, Liina Kinkar, Neil D. Young, Huimin Cai, Marshall W. Lightowlers, Charles Gauci, Abdul Jabbar, Bill C. H. Chang, Tao Wang, Andreas Hofmann, Anson V. Koehler, Junhua Li, Jiandong Li, Daxi Wang, Jiefang Yin, Huanming Yang, David J. Jenkins
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Raúl J. Bobes, Karel Estrada, Diana G. Rios-Valencia, Arturo Calderón-Gallegos, Patricia de la Torre, Julio C. Carrero, Alejandro Sanchez-Flores, Juan P. Laclette
Frontiers in Cellular and Infection Microbiology.2022;[Epub] CrossRef
Evolutionary Adaptations of Parasitic Flatworms to Different Oxygen Tensions
José de Jesús Martínez-González, Alberto Guevara-Flores, Irene Patricia del Arenal Mena
Antioxidants.2022; 11(6): 1102. CrossRef
Bioinformatics analysis and experimental verification of Notch signalling pathway-related miRNA–mRNA subnetwork in extracellular vesicles during Echinococcus granulosus encystation
Jian Gao, Xuan Zhou, Ling Liu, Guodong Lv, Qiulian Hou, Xiaofan Zhang, Yujuan Shen
Parasites & Vectors.2022;[Epub] CrossRef
iTRAQ-based comparative proteomic analysis in different developmental stages ofEchinococcus granulosus
Xin Li, Song Jiang, Xuhai Wang, Wenqiao Hui, Bin Jia
Parasite.2021; 28: 15. CrossRef
Genome-wide transcriptome analysis of the early developmental stages of Echinococcus granulosus protoscoleces reveals extensive alternative splicing events in the spliceosome pathway
Mohammad Ali Mohammadi, Majid Fasihi Harandi, Donald P. McManus, Mehdi Mansouri
Parasites & Vectors.2021;[Epub] CrossRef
Synergism therapeutic and immunoregulatory effects of Albendazole + rAd-mIL-28B against Echinococcosis in experiment-infected mice with protoscoleces
Yan Zhang, Jianghua Wang, Qingxia Yang, Zhi Li, Xiaoying Xu, Chong Chen, Zongjie Hou, Qi He, Li Sheng, Xingming Ma, Yanping Luo, Alessandra Morassutti
PLOS Neglected Tropical Diseases.2021; 15(11): e0009927. CrossRef

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Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae: Hae-Jin Sohn, Heekyoung Kang, Ga-Eun Seo, Jong-Hyun Kim, Suk-Yul Jung, Ho-Joon Shin; Korean J Parasitol 2017;55(3):233-238.
Published online June 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.3.233

Abstract
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Pathogenic Naegleria fowleri, Acanthamoeba castellanii, and Acanthamoeba polyphaga, are distributed worldwide. They are causative agents of primary amoebic meningoencephalitis or acanthamoebic keratitis in humans, respectively. Trophozoites encyst in unfavorable environments, such as exhausted food supply and desiccation. Until recently, the method of N. fowleri encystation used solid non-nutrient agar medium supplemented with heat-inactivated Escherichia coli; however, for the amoebic encystment of Acanthamoeba spp., a defined, slightly modified liquid media is used. In this study, in order to generate pure N. fowleri cysts, a liquid encystment medium (buffer 1) modified from Page’s amoeba saline was applied for encystation of N. fowleri. N. fowleri cysts were well induced after 24 hr with the above defined liquid encystment medium (buffer 1). This was confirmed by observation of a high expression of differential mRNA of nfa1 and actin genes in trophozoites. Thus, this liquid medium can replace the earlier non-nutrient agar medium for obtaining pure N. fowleri cysts. In addition, for cyst formation of Acanthamoeba spp., buffer 2 (adjusted to pH 9.0) was the more efficient medium. To summarize, these liquid encystment media may be useful for further studies which require axenic and pure amoebic cysts.

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Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts
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DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation: Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2017;55(2):115-120.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.115

Abstract
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Encystation mediating cyst specific cysteine proteinase (CSCP) of Acanthamoeba castellanii is expressed remarkably during encystation. However, the molecular mechanism involved in the regulation of CSCP gene expression remains unclear. In this study, we focused on epigenetic regulation of gene expression during encystation of Acanthamoeba. To evaluate methylation as a potential mechanism involved in the regulation of CSCP expression, we first investigated the correlation between promoter methylation status of CSCP gene and its expression. A 2,878 bp of promoter sequence of CSCP gene was amplified by PCR. Three CpG islands (island 1-3) were detected in this sequence using bioinformatics tools. Methylation of CpG island in trophozoites and cysts was measured by bisulfite sequence PCR. CSCP promoter methylation of CpG island 1 (1,633 bp) was found in 8.2% of trophozoites and 7.3% of cysts. Methylation of CpG island 2 (625 bp) was observed in 4.2% of trophozoites and 5.8% of cysts. Methylation of CpG island 3 (367 bp) in trophozoites and cysts was both 3.6%. These results suggest that DNA methylation system is present in CSCP gene expression of Acanthamoeba. In addition, the expression of encystation mediating CSCP is correlated with promoter CpG island 1 hypomethylation.

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DNA methylation modification: Dawn of research on cornea-related diseases
Quanhao Pan, Xiaoning Ge, Di Wang, Yuxi He
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Behroz Mahdavi Poor, Jalil Rashedi, Vahid Asgharzadeh, Amirali Mirmazhary, Nazila Gheitarani
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Acanthamoeba keratitis: new hopes for potential interventions for a curable but often refractory disease
Bader Saleem Alawfi, Naveed Ahmed Khan, David Lloyd, Ruqaiyyah Siddiqui
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Stimulation of Acanthamoeba castellanii excystment by enzyme treatment and consequences on trophozoite growth
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Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts
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The Korean Journal of Parasitology.2019; 57(3): 217. CrossRef

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Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii: Eun-Kyung Moon, Hyun-Hee Kong, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan; Korean J Parasitol 2017;55(2):109-114.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.109

Abstract
PDF

Protein arginine methyltransferase (PRMT) is an important epigenetic regulator in eukaryotic cells. During encystation, an essential process for Acanthamoeba survival, the expression of a lot of genes involved in the encystation process has to be regulated in order to be induced or inhibited. However, the regulation mechanism of these genes is yet unknown. In this study, the full-length 1,059 bp cDNA sequence of Acanthamoeba castellanii PRMT1 (AcPRMT1) was cloned for the first time. The AcPRMT1 protein comprised of 352 amino acids with a SAM-dependent methyltransferase PRMT-type domain. The expression level of AcPRMT1 was highly increased during encystation of A. castellanii. The EGFPAcPRMT1 fusion protein was distributed over the cytoplasm, but it was mainly localized in the nucleus of Acanthamoeba. Knock down of AcPRMT1 by synthetic siRNA with a complementary sequence failed to form mature cysts. These findings suggested that AcPRMT1 plays a critical role in the regulation of encystation of A. castellanii. The target gene of AcPRMT1 regulation and the detailed mechanisms need to be investigated by further studies.

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PRMT5 Inhibitor EPZ015666 Decreases the Viability and Encystment of Entamoeba invadens
Rigoberto Ortiz-Hernández, Elmer Joel Millán-Casarrubias, Jeni Bolaños, Susana Munguía-Robledo, Carlos Vázquez-Calzada, Elisa Azuara-Licéaga, Jesús Valdés, Mario Alberto Rodríguez
Molecules.2024; 30(1): 62. CrossRef
Comparative analysis of differentially expressed genes in Acanthamoeba after ingestion of Legionella pneumophila and Escherichia coli
Eun-Kyung Moon, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
Experimental Parasitology.2022; 232: 108188. CrossRef
Drug Discovery against Acanthamoeba Infections: Present Knowledge and Unmet Needs
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Free-living amoebae and squatters in the wild: ecological and molecular features
Ascel Samba-Louaka, Vincent Delafont, Marie-Hélène Rodier, Estelle Cateau, Yann Héchard
FEMS Microbiology Reviews.2019; 43(4): 415. CrossRef

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Identification of Protein Arginine Methyltransferase 5 as a Regulator for Encystation of Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong; Korean J Parasitol 2016;54(2):133-138.
Published online April 30, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.2.133

Abstract
PDF

Encystation is an essential process for Acanthamoeba survival under nutrient-limiting conditions and exposure to drugs. The expression of several genes has been observed to increase or decrease during encystation. Epigenetic processes involved in regulation of gene expression have been shown to play a role in several pathogenic parasites. In the present study, we identified the protein arginine methyltransferase 5 (PRMT5), a known epigenetic regulator, in Acanthamoeba castellanii. PRMT5 of A. castellanii (AcPRMT5) contained domains found in S-adenosylmethionine-dependent methyltransferases and in PRMT5 arginine-N-methyltransferase. Expression levels of AcPRMT5 were increased during encystation of A. castellanii. The EGFP-PRMT5 fusion protein was mainly localized in the nucleus of trophozoites. A. castellanii transfected with siRNA designed against AcPRMT5 failed to form mature cysts. The findings of this study lead to a better understanding of epigenetic mechanisms behind the regulation of encystation in cyst-forming pathogenic protozoa.

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Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher, Sébastien Pomel
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Oxidase enzyme genes are differentially expressed during Acanthamoeba castellanii encystment
Christian Q. Scheckhuber, Rebeca Damián Ferrara, Jesús Gómez-Montalvo, Sutherland K. Maciver, Alvaro de Obeso Fernández del Valle
Parasitology Research.2024;[Epub] CrossRef
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Bader Saleem Alawfi, Naveed Ahmed Khan, David Lloyd, Ruqaiyyah Siddiqui
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Biological characteristics and pathogenicity of Acanthamoeba
Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
Frontiers in Microbiology.2023;[Epub] CrossRef
Comparative analysis of differentially expressed genes in Acanthamoeba after ingestion of Legionella pneumophila and Escherichia coli
Eun-Kyung Moon, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
Experimental Parasitology.2022; 232: 108188. CrossRef
Stimulation of Acanthamoeba castellanii excystment by enzyme treatment and consequences on trophozoite growth
Zineb Fechtali-Moute, Philippe M. Loiseau, Sébastien Pomel
Frontiers in Cell and Developmental Biology.2022;[Epub] CrossRef
Free-living amoebae and squatters in the wild: ecological and molecular features
Ascel Samba-Louaka, Vincent Delafont, Marie-Hélène Rodier, Estelle Cateau, Yann Héchard
FEMS Microbiology Reviews.2019; 43(4): 415. CrossRef
Encystation: the most prevalent and underinvestigated differentiation pathway of eukaryotes
Pauline Schaap, Christina Schilde
Microbiology.2018; 164(5): 727. CrossRef
Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii
Eun-Kyung Moon, Hyun-Hee Kong, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan
The Korean Journal of Parasitology.2017; 55(2): 109. CrossRef
DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation
Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
The Korean Journal of Parasitology.2017; 55(2): 115. CrossRef

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Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong; Korean J Parasitol 2014;52(2):131-135.
Published online April 18, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.2.131

Abstract
PDF

Acanthamoeba cysts are resistant to unfavorable physiological conditions and various disinfectants. Acanthamoeba cysts have 2 walls containing various sugar moieties, and in particular, one third of the inner wall is composed of cellulose. In this study, it has been shown that down-regulation of cellulose synthase by small interfering RNA (siRNA) significantly inhibits the formation of mature Acanthamoeba castellanii cysts. Calcofluor white staining and transmission electron microscopy revealed that siRNA transfected amoeba failed to form an inner wall during encystation and thus are likely to be more vulnerable. In addition, the expression of xylose isomerase, which is involved in cyst wall formation, was not altered in cellulose synthase down-regulated amoeba, indicating that cellulose synthase is a crucial factor for inner wall formation by Acanthamoeba during encystation.

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Oxidase enzyme genes are differentially expressed during Acanthamoeba castellanii encystment
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Parasitology Research.2024;[Epub] CrossRef
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The role of the Acanthamoeba castellanii Sir2-like protein in the growth and encystation of Acanthamoeba
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DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation
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Identification of Atg8 Isoform in Encysting Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2013;51(5):497-502.
Published online October 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.5.497

Abstract
PDF

Autophagy-related protein 8 (Atg8) is an essential component of autophagy formation and encystment of cyst-forming parasites, and some protozoa, such as, Acanthamoeba, Entamoeba, and Dictyostelium, have been reported to possess a type of Atg8. In this study, an isoform of Atg8 was identified and characterized in Acanthamoeba castellanii (AcAtg8b). AcAtg8b protein was found to encode 132 amino acids and to be longer than AcAtg8 protein, which encoded 117 amino acids. Real-time PCR analysis showed high expression levels of AcAtg8b and AcAtg8 during encystation. Fluorescence microscopy demonstrated that AcAtg8b is involved in the formation of the autophagosomal membrane. Chemically synthesized siRNA against AcAtg8b reduced the encystation efficiency of Acanthamoeba, confirming that AcAtg8b, like AcAtg8, is an essential component of cyst formation in Acanthamoeba. Our findings suggest that Acanthamoeba has doubled the number of Atg8 gene copies to ensure the successful encystation for survival when 1 copy is lost. These 2 types of Atg8 identified in Acanthamoeba provide important information regarding autophagy formation, encystation mechanism, and survival of primitive, cyst-forming protozoan parasites.

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Brief Communication

Short-Cut Pathway to Synthesize Cellulose of Encysting Acanthamoeba: Eun-Kyung Moon, Hyun-Hee Kong; Korean J Parasitol 2012;50(4):361-364.
Published online November 26, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.4.361

Abstract
PDF

The mature cyst of Acanthamoeba is highly resistant to various antibiotics and therapeutic agents. Cyst wall of Acanthamoeba are composed of cellulose, acid-resistant proteins, lipids, and unidentified materials. Because cellulose is one of the primary components of the inner cyst wall, cellulose synthesis is essential to the process of cyst formation in Acanthamoeba. In this study, we hypothesized the key and short-step process in synthesis of cellulose from glycogen in encysting Acanthamoeba castellanii, and confirmed it by comparing the expression pattern of enzymes involving glycogenolysis and cellulose synthesis. The genes of 3 enzymes, glycogen phosphorylase, UDP-glucose pyrophosphorylase, and cellulose synthase, which are involved in the cellulose synthesis, were expressed high at the 1st and 2nd day of encystation. However, the phosphoglucomutase that facilitates the interconversion of glucose 1-phosphate and glucose 6-phosphate expressed low during encystation. This report identified the short-cut pathway of cellulose synthesis required for construction of the cyst wall during the encystation process in Acanthamoeba. This study provides important information to understand cyst wall formation in encysting Acanthamoeba.

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Inhibition of GABA metabolism by β-lactam antibiotics affects encystation in Acanthamoeba
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Oxidase enzyme genes are differentially expressed during Acanthamoeba castellanii encystment
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Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
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Zineb Fechtali-Moute, Philippe M. Loiseau, Sébastien Pomel
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The role of the Acanthamoeba castellanii Sir2-like protein in the growth and encystation of Acanthamoeba
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Parasites & Vectors.2020;[Epub] CrossRef
Encystation: the most prevalent and underinvestigated differentiation pathway of eukaryotes
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Microbiology.2018; 164(5): 727. CrossRef
Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba
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The Korean Journal of Parasitology.2014; 52(2): 131. CrossRef

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Original Articles

Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii: Eun-Kyung Moon, Ying-Hua Xuan, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong; Korean J Parasitol 2011;49(4):341-347.
Published online December 16, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.4.341

Abstract
PDF

Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba.

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The role of the Acanthamoeba castellanii Sir2-like protein in the growth and encystation of Acanthamoeba
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Soil protists: a fertile frontier in soil biology research
Stefan Geisen, Edward A D Mitchell, Sina Adl, Michael Bonkowski, Micah Dunthorn, Flemming Ekelund, Leonardo D Fernández, Alexandre Jousset, Valentyna Krashevska, David Singer, Frederick W Spiegel, Julia Walochnik, Enrique Lara
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DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation
Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
The Korean Journal of Parasitology.2017; 55(2): 115. CrossRef
Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii
Eun-Kyung Moon, Hyun-Hee Kong, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan
The Korean Journal of Parasitology.2017; 55(2): 109. CrossRef
Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii
Yu-Ran Lee, Byoung-Kuk Na, Eun-Kyung Moon, Su-Min Song, So-Young Joo, Hyun-Hee Kong, Youn-Kyoung Goo, Dong-Il Chung, Yeonchul Hong, Patricia Talamas-Rohana
PLOS ONE.2015; 10(6): e0129884. CrossRef
Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba
Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong
The Korean Journal of Parasitology.2014; 52(2): 131. CrossRef
Microarray and KOG analysis of Acanthamoeba healyi genes up-regulated by mouse-brain passage
Eun-Kyung Moon, Ying-Hua Xuan, Hyun-Hee Kong
Experimental Parasitology.2014; 143: 69. CrossRef
Proteomic profiling of the infective trophozoite stage of Acanthamoeba polyphaga
Karin Silva Caumo, Karina Mariante Monteiro, Thiely Rodrigues Ott, Vinicius José Maschio, Glauber Wagner, Henrique Bunselmeyer Ferreira, Marilise Brittes Rott
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Cysteine Protease Inhibitor (AcStefin) Is Required for Complete Cyst Formation of Acanthamoeba
Jung-Yub Lee, Su-Min Song, Eun-Kyung Moon, Yu-Ran Lee, Bijay Kumar Jha, Dinzouna-Boutamba Sylvatrie Danne, Hee-Jae Cha, Hak Sun Yu, Hyun-Hee Kong, Dong-Il Chung, Yeonchul Hong
Eukaryotic Cell.2013; 12(4): 567. CrossRef
Cysteine protease involving in autophagosomal degradation of mitochondria during encystation of Acanthamoeba
Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong
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Short-Cut Pathway to Synthesize Cellulose of Encysting Acanthamoeba
Eun-Kyung Moon, Hyun-Hee Kong
The Korean Journal of Parasitology.2012; 50(4): 361. CrossRef
Protein kinase C signaling molecules regulate encystation of Acanthamoeba
Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong
Experimental Parasitology.2012; 132(4): 524. CrossRef

9,628 View
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Atg3-Mediated Lipidation of Atg8 Is Involved in Encystation of Acanthamoeba: Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong; Korean J Parasitol 2011;49(2):103-108.
Published online June 14, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.2.103

Abstract
PDF

Autophagy is a catabolic process involved in the degradation of a cell's own components for cell growth, development, homeostasis, and the recycling of cellular products. Autophagosome is an essential component in the protozoan parasite during differentiation and encystation. The present study identified and characterized autophagy-related protein (Atg) 3, a member of Atg8 conjugation system, in Acanthamoeba castellanii (AcAtg3). AcAtg3 encoding a 304 amino acid protein showed high similarity with the catalytic cysteine site of other E2 like enzymes of ubiquitin system. Predicted 3D structure of AcAtg3 revealed a hammer-like shape, which is the characteristic structure of E2-like enzymes. The expression level of AcAtg3 did not increase during encystation. However, the formation of mature cysts was significantly reduced in Atg3-siRNA transfected cells in which the production of Atg8-phosphatidylethanolamine conjugate was inhibited. Fluorescent microscopic analysis revealed that dispersed AcAtg3-EGFP fusion protein gathered around autophagosomal membranes during encystation. These results provide important information for understanding autophagic machinery through the lipidation reaction mediated by Atg3 in Acanthamoeba.

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Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
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Ac-HSP20 regulates autophagy and promotes the encystation of Acanthamoeba castellanii by inhibiting the PI3K/AKT/mTOR signaling pathway
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Biological characteristics and pathogenicity of Acanthamoeba
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Phragmites australis (Cav.) Trin. ex Steud. Extract Induces Apoptosis-like Programmed Cell Death in Acanthamoeba castellanii Trophozoites
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Transcriptomic changes across the life cycle of Trypanosoma cruzi II
Lissa Cruz-Saavedra, Gustavo A. Vallejo, Felipe Guhl, Juan David Ramírez
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The role of the Acanthamoeba castellanii Sir2-like protein in the growth and encystation of Acanthamoeba
So-Young Joo, Ja Moon Aung, Minsang Shin, Eun-Kyung Moon, Hyun-Hee Kong, Youn-Kyoung Goo, Dong-Il Chung, Yeonchul Hong
Parasites & Vectors.2020;[Epub] CrossRef
Apoptosis of Acanthamoeba castellanii Trophozoites Induced by Oleic Acid
Duo Wu, Ke Qiao, Meng Feng, Yongfeng Fu, Junlong Cai, Yihong Deng, Hiroshi Tachibana, Xunjia Cheng
Journal of Eukaryotic Microbiology.2018; 65(2): 191. CrossRef
Identification and ultrastructural characterization of Acanthamoeba bacterial endocytobionts belonging to the Alphaproteobacteria class
Li Li Chan, Joon Wah Mak, Stephen Ambu, Pei Yee Chong, Lorenzo Brusetti
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Atg8 is involved in endosomal and phagosomal acidification in the parasitic protist E ntamoeba histolytica
Karina Picazarri, Kumiko Nakada‐Tsukui, Kumiko Tsuboi, Eri Miyamoto, Naoko Watanabe, Eiryo Kawakami, Tomoyoshi Nozaki
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Experimental Parasitology.2015; 159: 46. CrossRef
Autophagy Inhibitors as a Potential Antiamoebic Treatment for Acanthamoeba Keratitis
Eun-Kyung Moon, So-Hee Kim, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong
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Chloroquine Has a Cytotoxic Effect on Acanthamoeba Encystation through Modulation of Autophagy
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Antimicrobial Agents and Chemotherapy.2014; 58(10): 6235. CrossRef
Identification of Atg8 Isoform in Encysting Acanthamoeba
Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong
The Korean Journal of Parasitology.2013; 51(5): 497. CrossRef
Cysteine protease involving in autophagosomal degradation of mitochondria during encystation of Acanthamoeba
Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong
Molecular and Biochemical Parasitology.2012; 185(2): 121. CrossRef
Protein kinase C signaling molecules regulate encystation of Acanthamoeba
Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong
Experimental Parasitology.2012; 132(4): 524. CrossRef
Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites ofAcanthamoeba castellanii
Eun-Kyung Moon, Ying-Hua Xuan, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong
The Korean Journal of Parasitology.2011; 49(4): 341. CrossRef

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Construction of EST Database for Comparative Gene Studies of Acanthamoeba: Eun-Kyung Moon, Joung-Ok Kim, Ying-Hua Xuan, Young-Sun Yun, Se Won Kang, Yong Seok Lee, Tae-In Ahn, Yeon-Chul Hong, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2009;47(2):103-107.
Published online May 26, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.2.103

Abstract
PDF

The genus Acanthamoeba can cause severe infections such as granulomatous amebic encephalitis and amebic keratitis in humans. However, little genomic information of Acanthamoeba has been reported. Here, we constructed Acanthamoeba expressed sequence tags (EST) database (Acanthamoeba EST DB) derived from our 4 kinds of Acanthamoeba cDNA library. The Acanthamoeba EST DB contains 3,897 EST generated from amebae under various conditions of long term in vitro culture, mouse brain passage, or encystation, and downloaded data of Acanthamoeba from National Center for Biotechnology Information (NCBI) and Taxonomically Broad EST Database (TBestDB). The almost reported cDNA/genomic sequences of Acanthamoeba provide stand alone BLAST system with nucleotide (BLAST NT) and amino acid (BLAST AA) sequence database. In BLAST results, each gene links for the significant information including sequence data, gene orthology annotations, relevant references, and a BlastX result. This is the first attempt for construction of Acanthamoeba database with genes expressed in diverse conditions. These data were integrated into a database (http://www.amoeba.or.kr).

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Protein kinase C signaling molecules regulate encystation of Acanthamoeba
Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong
Experimental Parasitology.2012; 132(4): 524. CrossRef
Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites ofAcanthamoeba castellanii
Eun-Kyung Moon, Ying-Hua Xuan, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong
The Korean Journal of Parasitology.2011; 49(4): 341. CrossRef
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Brief Communication

Differentially expressed genes of Acanthamoeba castellanii during encystation: Eun-Kyung Moon, Dong-Il Chung, Yeon-Chul Hong, Hyun-Hee Kong; Korean J Parasitol 2007;45(4):283-285.
Published online December 20, 2007; DOI: https://doi.org/10.3347/kjp.2007.45.4.283

Abstract
PDF

To examine the expressed gene profile during encystation of Acanthamoeba castellanii Castellani, we used differentially expressed gene (DGE) screening by RT-PCR with 20 sets of random primers. From this analysis, we found that approximately 16 genes showed upregulation during encystation. We chose 6 genes, which had relatively higher expression levels, for further investigation. Based on homology search in database, DEG2 showed 55% of similarity with xylose isomerase, DEG9 showed 37% of similarity with Na P-type ATPase, and DEG14 showed 77% of similarity with subtilisin-like serine proteinase. DEG3 and DEG26 were identified as hypothetical proteins and DEG25 exhibited no significant similarity to any known protein. Encystation of Acanthamoeba has been suggested to be a process to resist adverse environmental or nutritional conditions. Further characterization studies of these genes may provide us with more information on the encystation mechanism of Acanthamoeba.

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Oxidase enzyme genes are differentially expressed during Acanthamoeba castellanii encystment
Christian Q. Scheckhuber, Rebeca Damián Ferrara, Jesús Gómez-Montalvo, Sutherland K. Maciver, Alvaro de Obeso Fernández del Valle
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Fernando Pérez-Cota, Richard J. Smith, Hany M. Elsheikha, Matt Clark
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Encystation: the most prevalent and underinvestigated differentiation pathway of eukaryotes
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Ludmila Karen dos Santos Silva, Paulo Victor Miranda Boratto, Bernard La Scola, Cláudio Antônio Bonjardim, Jônatas Santos Abrahão
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Autophagy protein 12 plays an essential role in Acanthamoeba encystation
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Experimental Parasitology.2015; 159: 46. CrossRef
Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba
Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong
The Korean Journal of Parasitology.2014; 52(2): 131. CrossRef
Chloroquine Has a Cytotoxic Effect on Acanthamoeba Encystation through Modulation of Autophagy
Bijay Kumar Jha, Hui-Jung Jung, Incheol Seo, Hyun Ah Kim, Seong-Il Suh, Min-Ho Suh, Won-Ki Baek
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Original Article

In vivo determination of the gap2 gene promoter activity in Giardia lamblia: Hye-Won Yang, Juri Kim, Tai-Soon Yong, Soon-Jung Park; Korean J Parasitol 2006;44(1):21-26.
Published online March 20, 2006; DOI: https://doi.org/10.3347/kjp.2006.44.1.21

Abstract
PDF

A shuttle vector for Escherichia coli and Giardia lamblia was modified to produce a reporter plasmid, which monitors the expression of prescribed gene in G. lamblia by measuring its luciferase activity. Promoter regions of the gap2 gene, one of the genes induced during encystation, were cloned into this plasmid, and the resultant constructs were then transfected into trophozoites of G. lamblia. Transgenic trophozoites containing one of the 3 gap2-luc reporters were induced to encystation, and characterized with respect to gap2 gene expression by measuring their luciferase activities. Giardia containing a gap2-luc fusion of 112-bp upstream region showed full induction of luciferase activity during encystation.

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