지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2025.2
- 수록면
- 25 - 36 (12page)
이용수
초록· 키워드
오류제보하기
Acanthamoeba is an opportunistic pathogen responsible for granulomatous amoebic encephalitis and amoebic keratitis. Despite its clinical significance, effective treatments remain challenging due to a limited understanding of its pathogenic mechanism. This study developed a genetic manipulation system in Acanthamoeba to facilitate gene function and drug screening studies. We applied the Cre/loxP system to integrate the gene encoding the tdTomato fluorescent protein into the genome of Acanthamoeba castellanii via homologous recombination. The polyubiquitin gene and its untranslated regions were identified and verified, after which the tdTomato gene was cloned between the untranslated regions of the polyubiquitin gene. The construct was then introduced into the Acanthamoeba genome using a modified pLPBLP vector containing loxP sites.
Cre recombinase was utilized to remove the neomycin resistance cassette flanked by loxP sites, and genetically modified cells were selected by clonal dilution. The integration of the tdTomato gene, confirmed through PCR and fluorescence microscopy, showed stable expression in both trophozoites and cysts without the need for antibiotic selection. We demonstrated the feasibility of antibiotic-free reporter gene expression in Acanthamoeba. The system provides a valuable tool for functional genomics, allowing us to explore gene functions in Acanthamoeba and develop reliable drug screening models. Furthermore, the ability to express genes without the continuous use of selection markers opens up new possibilities for studying the pathobiology of this pathogen and advancing the development of novel therapeutic strategies against Acanthamoeba infections.
Cre recombinase was utilized to remove the neomycin resistance cassette flanked by loxP sites, and genetically modified cells were selected by clonal dilution. The integration of the tdTomato gene, confirmed through PCR and fluorescence microscopy, showed stable expression in both trophozoites and cysts without the need for antibiotic selection. We demonstrated the feasibility of antibiotic-free reporter gene expression in Acanthamoeba. The system provides a valuable tool for functional genomics, allowing us to explore gene functions in Acanthamoeba and develop reliable drug screening models. Furthermore, the ability to express genes without the continuous use of selection markers opens up new possibilities for studying the pathobiology of this pathogen and advancing the development of novel therapeutic strategies against Acanthamoeba infections.
목차
Abstract
Introduction
Materials and Methods
Results
Discussion
References
참고문헌 (0)
참고문헌 신청함께 읽어보면 좋을 논문
논문 유사도에 따라 DBpia 가 추천하는 논문입니다. 함께 보면 좋을 연관 논문을 확인해보세요!
-
Acanthamoeba in Southeast Asia – Overview and Challenges
Parasites, Hosts and Diseases
2019 .08
-
Can Acanthamoeba keratitis be properly diagnosed without culture in the real-world clinical microbiology laboratory?: a case report
Annals of Clinical Microbiology
2024 .06
-
Identification and Genotypic Characterization of Potentially Pathogenic Acanthamoeba Isolated from Tap Water in Wuxi, China
Parasites, Hosts and Diseases
2018 .12
-
Isolation and Genotyping of Acanthamoeba spp. as Neglected Parasites in North of Iran
Parasites, Hosts and Diseases
2016 .08
-
Identification of essential genes for Acanthamoeba castellanii excystation during encystation and excystation
Parasites, Hosts and Diseases
2024 .11
-
The First Acanthamoeba keratitis Case of Non-Contact Lens Wearer with HIV Infection in Thailand
Parasites, Hosts and Diseases
2019 .10
-
DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation
Parasites, Hosts and Diseases
2017 .04
-
Comparison of Proteins Secreted into Extracellular Space of Pathogenic and Non-pathogenic Acanthamoeba castellanii
Parasites, Hosts and Diseases
2018 .12
-
Characterization of a Peptide Antibody Specific to the Adenylyl Cyclase-Associated Protein of Acanthamoeba castellanii
Parasites, Hosts and Diseases
2022 .02
-
Sirtinol Supresses Trophozoites Proliferation and Encystation of Acanthamoeba via Inhibition of Sirtuin Family Protein
Parasites, Hosts and Diseases
2022 .02
-
Epigenetic Regulation of Gene Structure and Function with Improved Cell-Permeable Cre (iCP-Cre) Recombinase for Site-Specific Recombination in Genetically Engineered Animals
한국실험동물학회 학술발표대회 논문집
2016 .08
-
Phagocytosis-associated genes in Acanthamoeba castellanii feeding on Escherichia coli
Parasites, Hosts and Diseases
2023 .11
-
Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae
Parasites, Hosts and Diseases
2017 .06
-
Molecular and Biochemical Properties of a Cysteine Protease of Acanthamoeba castellanii
Parasites, Hosts and Diseases
2018 .10
-
Efficient Production of loxP Knock-in Mouse using CRISPR/Cas9 System
대한의생명과학회지
2020 .06
-
Differentially Expressed Gene Profile of Acanthamoeba castellanii Induced by an Endosymbiont Legionella pneumophila
Parasites, Hosts and Diseases
2021 .02
-
Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii
Parasites, Hosts and Diseases
2017 .04
-
Efficacy of Korean Multipurpose Contact Lens Disinfecting Solutions against Acanthamoeba castellanii
Parasites, Hosts and Diseases
2016 .12
-
Identification of Protein Arginine Methyltransferase 5 as a Regulator for Encystation of Acanthamoeba
Parasites, Hosts and Diseases
2016 .04
-
Specific Detection of Acanthamoeba species using Polyclonal Peptide Antibody Targeting the Periplasmic Binding Protein of A. castellanii
Parasites, Hosts and Diseases
2022 .04
이 논문의 저자 정보
최근 본 자료
댓글(0)
0
UCI(KEPA) : I410-151-25-02-092594467