Molecular characterization and expressional analysis of two poly (ADP-ribose) polymerase (PARP) domain-containing Gig2 isoforms in rockfish (Sebastes schlegelii) and their antiviral activity against viral hemorrhagic septicemia virus
K.P. Madushani, K.A.S.N. Shanaka, Rajamanthrilage Kasun Madusanka, Jehee Lee
언어
영어(ENG)
URL
https://www.earticle.net/Article/A406958
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4,000원
원문정보
초록
영어
Remedies toward sustainable aquaculture rely upon research that unveils the molecular mechanisms behind host immunity and their interactions with pathogens. Antiviral defense is a major innate immune response in fish. The antiviral protein GCHV-induced gene-2 (Gig2), a member of the interferon-stimulated gene (ISG), was identified and characterized from rockfish (Sebastes schlegelii). Gig2 exists in two isoforms, namely, SsGig2-I1 and SsGig2-I2, in rockfish with lengths of 163 and 223 bp, respectively. Bioinformatic analysis indicated the availability of poly (ADP-ribose) polymerase domain in both proteins, and 51.3% identity and 71.3% similarity between both isoforms were observed. The basal expression pattern revealed the highest tissue-specific expression in rockfish gills for both isoforms. The immune challenge experiment disclosed a distinctive and strong expression of each transcript in the presence of poly I:C. Both isoforms are localized in the endoplasmic reticulum. Interferon (IFN) pathway gene analysis revealed no significant upregulation of IFN related genes. Viral hemorrhagic septicemia virus (VHSV) gene expression analysis revealed strong downregulation of viral transcripts after 48 h of infection in the presence of Gig2 isoforms. Collectively, these results indicate the protective role of Gig2 in rockfish against VHSV infection and help broaden our understanding of the innate immunity of fish.
목차
ABSTRACT 1. Introduction 2. Materials and methods 2.1. Identification of Gig2 from rockfish 2.2. Rearing fish 2.3. Tissue sampling and the immune challenge experiment 2.4. Construction of the expression plasmid 2.5. Cell culture and transfection 2.6. Downstream gene expressions 2.7. Viral hemorrhagic septicemia virus (VHSV) gene expression 2.8. RNA extraction and cDNA synthesis 2.9. Quantitative real-time PCR (qPCR) 2.10. Sub-cellular localization 2.11. Statistical analysis 3. Results and discussion 3.1. In silico analysis 3.2. Distribution of Gig2 in tissues 3.3. Immune challenge experiment 3.4. Subcellular localization of Gig2 3.5. The function of PARP domain in downstream gene expression 3.6. VHSV replication in the presence of SsGig2-I1 or SsGig2-I2 4. Conclusion References
K.P. Madushani [ Department of Marine Life Sciences, Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
K.A.S.N. Shanaka [ Department of Marine Life Sciences, Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
Rajamanthrilage Kasun Madusanka [ Department of Marine Life Sciences, Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea ]
Jehee Lee [ Department of Marine Life Sciences, Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
Corresponding Author
제주대학교 해양과학연구소 [Marine Science Institute Jeju National University]
설립연도
1968
분야
농수해양>수산학
소개
해양의 실체와 그 자원의 합리적 이용과 관리방법을 규명하기 위하여 해양물리, 화학, 생물, 지질, 공학등 여러 관련분야에 관한 다양한 연구를 수행하며, 환경의 보전과 보호를 위해서 수질, 대기, 상·하수도, 폐기물, 사회적 환경 등의 연구를 통하여 쾌적하고 청정한 환경을 보전하기 위한 다양한 연구를 수행하고, 또한 해양 및 환경분야의 우수한 인력양성, 국내외 학술교류의 증진, 학·산 협동체제의 확립 등을 통하여 해양과 환경에 관한 기초 및 응용적 학술연구를 종합적으로 수행함을 목적으로 한다.
간행물
간행물명
해양과학연구소 연구논문집 [Bulletin of the Marine Science Institute]