Functional Evaluation of the Rockbream (Oplegnathus fasciatus) Beta-actin Promoter as a Candidate Regulatory Element for DNA Vaccination
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
The potential utility of the rockbream (Oplegnathus fasciatus) β-actin 5´-upstream sequence as a regulatory element for DNA vaccination was evaluated based on in vitro and in vivo heterologous expression assays. In the in vitro transfection experiment, the efficacy of the rockbream β-actin promoter to drive the expression of a downstream lacZ gene was significantly higher (more than fourfold) than that of the human cytomegalovirus (hCMV) promoter in two fish cell lines (grunt Haemulon plumierii fin and bluegill Lepomis macrochirus fry cell lines). In contrast, the functional activity of the rockbream β-actin promoter was hardly detectable in a mammalian mouse embryonic fibroblast cell line. Rockbream skeletal muscles injected in vivo with a GFP reporter construct driven by the β-actin promoter displayed the significantly higher expression of a GFP protein (more than threefold) than did those injected with hCMV promoter driven construct. Data from this study suggest that the homologous rockbream β-actin promoter could be used as a potential regulator for DNA vaccination in this species.
Abstract
The potential utility of the rockbream (Oplegnathus fasciatus) β-actin 5´-upstream sequence as a regulatory element for DNA vaccination was evaluated based on in vitro and in vivo heterologous expression assays. In the in vitro transfection experiment, the efficacy of the rockbream β-actin promoter to drive the expression of a downstream lacZ gene was significantly higher (more than fourfold) than that of the human cytomegalovirus (hCMV) promoter in two fish cell lines (grunt Haemulon plumierii fin and bluegill Lepomis macrochirus fry cell lines). In contrast, the functional activity of the rockbream β-actin promoter was hardly detectable in a mammalian mouse embryonic fibroblast cell line. Rockbream skeletal muscles injected in vivo with a GFP reporter construct driven by the β-actin promoter displayed the significantly higher expression of a GFP protein (more than threefold) than did those injected with hCMV promoter driven construct. Data from this study suggest that the homologous rockbream β-actin promoter could be used as a potential regulator for DNA vaccination in this species.
참고문헌 (21)
- Adams, A, "Biotechnology offers revolution to fish health management", Trends. Bio-tech., 24, pp.201~205 (2006)
- Chan, W.K, "Polymerase chain reaction amplification and functional characterization of sockeye salmon histone H3, metallothionein-B and protamine promoters", Mol. Mar. Biol. Biotechnol., 2, pp.308~318 (1993)
- Choi, S.K., "Organ distribution of red sea bream iridovirus (RSIV) DNA in asymptomatic yearling and fingerling rock bream (Oplegnathus fasciatus) and effects of water temperature on transition of RSIV into acute phase", Aquaculture, (256), pp.23~26 (2006)
- Garver, "Efficacy of an infectious hematopoietic necrosis (IHN) virus DNA vaccine in Chinook Oncorhynchus tshawytscha and sockeye O. nerka salmon", Diseases of aquatic organisms, 64, pp.13~22 (2005)
- Gómez-Chiarii, M, "Evaluation of eukaryotic promoters for the construction of DNA vaccines for aquaculture", Genet. Anal. Biomol. Eng., 15, pp.121~124 (1999)
- Hackett, P.B, "The molecular genetics of transgenic fish. In: Recent Advances in Marine Biotechnology", pp.77~145 (2000), Science Publishers Inc.
- Heppell, J, "Application of DNA vaccine technology to aquaculture", Adv. Drug Del. Rev., 43, pp.29~43 (2000)
- Hwang, G.L., "Isolation and characterization of tilapia β-actin promoter and comparison of its activity with carp β-actin promoter", Biochim. Biophys. Acta, (1125), pp.11~18 (2003)
- Lee, S.Y., "Molecular characterization of rockbream (Oplegnathus fasciatus) cytoskeletal β-actin gene and 5´-upstream flanking region", J. Aquat. Fish. Sci., ~ (2009)
- Lorenzen, N, "DNA vaccines for aquacultured fish", Rev. Sci. Tech., 24, pp.201~213 (2005)
- Mikalsen, A.B., "Protective effects of a DNA vaccine expressing the infectious salmon anemia virus hemagglutinin-esterase in Atlantic salmon", Vaccine, 23, pp.4895~4905 (2005)
- Nam, Y.K., "Autotransgenic and allotransgenic manipulation of growth traits in fish for aquaculture", J. Fish Biol., 72, pp.1~26 (2008)
- Nam, Y.K., "Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach Misgurnus mizolepis", Transgenic Res., 10, pp.353~362 (2001)
- Noh, J.K., "Genomic cloning of mud loach Misgurnus mizolepis (Cypriniformes, Cobiti-dae) β-actin gene and usefulness of its promoter region for fish transgenesis", Mar. Biotechnol., 5, pp.244~252 (2003)
- Pasnik, D.J, "Immunogenic and protective effects of a DNA vaccine for Mycobac-terium marinum in fish", Vet. Immunol. Immuno-phathol., (103), pp.195~206 (2005)
- Reece, K.S., "Function and evolution of actins", Evol. Biol., 26, pp.1~34 (1992)
- Robertson, J.S, "Assuring the quality, safety, and efficacy of DNA vaccines", Methods Mol. Med., (127), pp.363~374 (2006)
- Ruiz, S., "In vitro search for alternative promoters to the human immediate early cytomegalovirus (IE-CMV) to express the G gene of viral haemorrhagic septicemia virus (VHSV) in fish epithelial cells", Vaccine, 26, pp.6620~6629 (2008)
- Sanchez, E., "Expression of inducible CC chemokines in rainbow trout (Oncor-hynchus mykiss) in response to a viral heamorrhagic septicemia virus (VHSV) DNA vaccine and inter-leukin 8", Dev. Comp. Immunol., 31, pp.916~926 (2007)
- Verri, T., "Assessment of DNA vaccine potential for gilthead sea bream (Sparus aurata) by intra-muscular injection of reporter gene", Fish Shellfish Immunol., 15, pp.283~295 (2003)
- Winkler, C., "Analysis of heterologous and homologous promoters and enhancers in vitro and in vivo by gene transfer into Japanese medaka (Oryzias latipes) and Xiphop-horus", Mol. Mar. Biol. Biotechnol., 1, pp.326~327 (1992)