수온변화에 따른 붉바리(Epinephelus akaara)의 heat shock protein (Hsp) 70 mRNA 발현

국문 초록 (Abstract)

한국의 고급 양식대상 어종인 붉바리(Epinephelus akaara)로부터 새로운 heat shock protein (Hsp) 70을 동정하였다. 붉바리 Hsp70 (RgHsp70)의 cDNA는 RACE (rapid amplification of cDNA ends)법을 사용하였고, RgHsp70 cDNA의 전장은 2,152 bp이고, 5’-terminal untranslated region (UTR)은 105 bp, 3’-terminal UTR은 274 bp, 590개의 아미노산을 암호화하는 open reading frame (ORF)는 1,773 bp였으며, 분자무게(molecular weight)는 64.9 kDa 및 등전위값(isoelectric point, pI)은 5.2였다. 추정되는 아미노산 비교 및 계통발생학적 분석 결과, 다른 어종과 마찬가지로 Hsp70 고유의 signature를 포함하는 것을 비롯하여 높은 유사성을 나타내었으므로 RgHsp70이 Hsp70 family임을 확인할 수 있었다 RgHsp70 mRNA는 간과 두신 조직에서 높은 발현을 보였으며, 48시간 동안 수온별(21, 18, 15 및 12℃) 노출후 간 조직에서 대조구인 21℃보다 12℃에서 발현이 증가함을 확인하였다. 본 연구에서는, 수온이 하강함에 따라 RgHsp70 mRNA 발현에 주요한 영향을 미치는 것으로 보아, 수온변화에 따른 스트레스로 인해 발현의 변화를 나타내는 주요 스트레스성 단백질임을 확인할 수 있었다.

다국어 초록 (Multilingual Abstract)

A new heat shock protein 70 was identified in red-spotted grouper (Epinephelus akaara) based on an expression analysis. The cDNA of red-spotted grouper Hsp70 (designated RgHsp70) was cloned by the rapid amplification of cDNA ends (RACE) techniques. The full-length of RgHsp70 cDNA was 2,152 bp, consisting of a 5’-terminal untranslated region (UTR) of 105 bp, a 3’-terminal UTR of 274 bp, and an open reading frame (ORF) of 1,773 bp that encode a polypeptide of 590 amino acids with a theoretical molecular weight of 64.9 kDa and an estimated isoelectric point of 5.2. Multiple alignment and phylogenetic analyses revealed that the RgHsp70 gene shares a high similarity with other Hsp70 fish genes. RgHsp70 contained all three classical Hsp70 family signatures. The results indicated the RgHsp70 is a member of the heat shock protein 70 family. RgHsp70 mRNA was predominately expressed in the liver, with reduced expression noted in the head-kidney tissues. The expression analysis of different water temperatures (21, 18, 15 and 12℃) for sampled livers revealed that expression gradually increased at 12℃ compared to 21℃. In this study, the effects of water temperature lowering on the physiological conditions were investigated, and the results revealed that novel RgHsp70 may be an important molecule involved in stress responses.

목차 (Table of Contents)

• 서론
• 재료 및 방법
• 결과 및 고찰
• References
• 초록

참고문헌 (Reference)

1 Ali, K. S., "Tissue-and stressor-specific differential expression of two hsc70 genes in carp" 307 : 503-509, 2003

2 Sharp, G. J. E., "The role of reactive oxygen species in the killing of the bacterial fish pathogen Aeromonas salmonicida by rainbow trout macrophages" 3 : 119-129, 1993

3 Yenari, M. A., "The neuroprotective potential of heat shock protein 70 (HSP70)" 5 : 525-531, 1999

4 Lindquist, S., "The heat-shock response" 55 : 1151-1191, 1986

5 Srivastava, P., "Roles of heat-shock proteins in innate and adaptive immunity" 2 : 185-194, 2002

6 Morimoto, R. I., "Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators" 12 : 3788-3796, 1998

7 Ojima, N., "Quantitative mRNA expression profiling of heat-shock protein families in rainbow trout" 329 : 51-57, 2005

8 Ming, J., "Molecular cloning and expression of two HSP70 genes in the Wuchang bream (Megalobrama amblycephala Yih)" 28 : 407-418, 2010

9 Konstantina, M., "Molecular cloning and characterization of two homologues of mannose-binding lectin in rainbow trout" 21 : 305-314, 2006

10 Zhang, X., "Molecular characterization of heat shock protein 70 gene transcripts during Vibrio harvei infection of humphead snapper, Lutjanus sanguineus" 37 : 897-910, 2011

11 Feder, M. E., "Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology" 61 : 243-282, 1999

12 Hofmann, G., "Heat-shock protein expression is absent in the Antartic fish Trematomus bernacchii (Family nototheniidae)" 203 : 2331-2339, 2000

13 Basu, N., "Heat shock proteins genes and their functional significance in fish" 295 : 173-183, 2002

14 Milani, V., "Heat shock protein 70: role in antigen presentation and immune stimulation" 18 : 563-575, 2002

15 Logue, J., "Heat injury and resistance adaptation in fish" 20 : 191-197, 1995

16 Deane, E. E., "Growth hormone increases hsc70/hsp70 expression and protects against apotosis in whole blood preparations from silver sea bream" 1040 : 288-292, 2005

17 Song, L., "Genome-wide identification of Hsp70 genes in channel catfish and their regulated expression after bacterial infection" 49 : 154-162, 2016

18 Zmijewski, M. A., "Functional similarities and differences of an archaeal HSP70 (DnaK) stress protein compared with its homologue from the bacterium Escherichia coli" 336 : 539-549, 2004

19 Chen, J., "F-type lectin involved in defense against bacterial infection in the pearl oyster (Pinctada martesii)" 30 : 750-754, 2011

20 Park, H., "Expression of heat shock protein 70 in the thermally stressed Antarctic clam Laternula elliptica" 12 : 275-282, 2007

21 Robert, J., "Evolution of heat shock pretein and immunity" 27 : 449-464, 2004

22 Deane, E. E., "Effects of GH, prolactin and cortisol on hepatic heat shock protein 70 expression in a marine teleost Sparus sarba" 161 : 413-421, 1999

23 Holmskov, U., "Collectins and ficolins: Humoral lectins of the innate immune defense" 21 : 547-578, 2003

24 Wang, Z., "Cloning and expression of heat shock protein 70 gene in the haemocytes of pearl oyster (Pinctada fucata, Gould 1850) responding to bacterial challenge" 26 : 639-645, 2009

25 Deane, E. E., "Cloning and characterization of the hsp70 multi-gene family from silver sea bream: Modulated gene expression between warm and cold temperature acclimation" 330 : 776-783, 2005

26 Graser, R. T., "Cloning and characterization of a 70kDa heat shock cognate (hsc70) gene from the zebra fish (Danio rerio)" 98 : 273-276, 1996

27 Listinsky, J. J., "Alpha-L-fucose: a potentially critical molecule in pathologic processes including neoplasia" 110 : 425-440, 1998