Molecular cloning, characterization and expression patterns of HSP70 and HSP60 in the pufferfish (Takifugu rubripes)

Takifugu rubripes is a commercially important fish in the Chinese aquaculture industry due to its excellent meat quality. In eukaryotic cells, heat shock proteins (HSPs) play a crucial role in maintaining intracellular protein homeostasis. They play significant roles in response to adverse environme...

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Main Authors: Kun-peng Fan, Ya-fang Liu, Zhi-qiang Zhang, Ying Liu, Peng-fei Liu
Format: Article
Language:English
Published: Elsevier 2021-11-01
Series:Aquaculture Reports
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2352513421002817
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author Kun-peng Fan
Ya-fang Liu
Zhi-qiang Zhang
Ying Liu
Peng-fei Liu
author_facet Kun-peng Fan
Ya-fang Liu
Zhi-qiang Zhang
Ying Liu
Peng-fei Liu
author_sort Kun-peng Fan
collection DOAJ
description Takifugu rubripes is a commercially important fish in the Chinese aquaculture industry due to its excellent meat quality. In eukaryotic cells, heat shock proteins (HSPs) play a crucial role in maintaining intracellular protein homeostasis. They play significant roles in response to adverse environmental stresses in aquatic animals. In this study, we used bioinformatics to comprehensively compare two different full-length cDNA sequences at the molecular level in the HSP family of T. rubripes. We performed various physicochemical studies such as race cloning and RT-qPCR, and found that TrHSP60 (575 amino acids) has a larger polypeptide, followed by TrHSP70–13 (441 amino acids). Homology and phylogenetic studies showed that these sequences have the greatest homology to Takifugu bimaculatus. The mRNA of the two genes were constitutively expressed in all tested tissues of untreated T. rubripes, including brain, muscle, kidney, liver, spleen, gill, and intestine, with highest expression levels in the intestine. Three experimental groups were established in this study, the H1 group (20 °C), H2 group (24 °C) and H3 group (28 °C), in order to analyze the expressed levels of TrHSP70 in T. rubripes. To understand the role of HSPs in temperature stress in T. rubripes, we analyzed the expression levels of HSPs in T. rubripes at different temperatures. The results showed that the mRNA expression of the HSP gene were influenced by temperature stress. Gene expression of TrHSP70 and TrHSP60 were mainly up-regulated at 12 h and 48 h in the H1 group, respectively. Meanwhile, gene expression of TrHSP70 was highest at 48 h in the H3 group and TrHSP60 reached was lowest at 24 h in the H3 group. The results showed that the two genes were constitutively expressed genes, which play an important role in maintaining normal physiological function and stress responses. The results show that TrHSP70 and TrHSP60 are constitutively expressed genes, which plays important role in maintaining normal physiological function and coping with stress.
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spelling doaj.art-8a059406873d403990f2fa684e52d8012022-12-21T18:14:01ZengElsevierAquaculture Reports2352-51342021-11-0121100865Molecular cloning, characterization and expression patterns of HSP70 and HSP60 in the pufferfish (Takifugu rubripes)Kun-peng Fan0Ya-fang Liu1Zhi-qiang Zhang2Ying Liu3Peng-fei Liu4College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China; Key Laboratory of Environment Controlled Aquaculture (KLECA), Ministry of Education, 52 Heishijiao Street, Dalian 116023, ChinaKey Laboratory of Environment Controlled Aquaculture (KLECA), Ministry of Education, 52 Heishijiao Street, Dalian 116023, China; College of Marine Technology and Environment, Dalian Ocean University, Dalian 116023, ChinaCollege of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China; Key Laboratory of Environment Controlled Aquaculture (KLECA), Ministry of Education, 52 Heishijiao Street, Dalian 116023, ChinaKey Laboratory of Environment Controlled Aquaculture (KLECA), Ministry of Education, 52 Heishijiao Street, Dalian 116023, China; College of Marine Technology and Environment, Dalian Ocean University, Dalian 116023, China; Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, ChinaKey Laboratory of Environment Controlled Aquaculture (KLECA), Ministry of Education, 52 Heishijiao Street, Dalian 116023, China; College of Marine Technology and Environment, Dalian Ocean University, Dalian 116023, China; Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China; Corresponding author at: College of Marine Technology and Environment, Dalian Ocean University, No. 52, Heishijiao Road, Dalian 116023, China.Takifugu rubripes is a commercially important fish in the Chinese aquaculture industry due to its excellent meat quality. In eukaryotic cells, heat shock proteins (HSPs) play a crucial role in maintaining intracellular protein homeostasis. They play significant roles in response to adverse environmental stresses in aquatic animals. In this study, we used bioinformatics to comprehensively compare two different full-length cDNA sequences at the molecular level in the HSP family of T. rubripes. We performed various physicochemical studies such as race cloning and RT-qPCR, and found that TrHSP60 (575 amino acids) has a larger polypeptide, followed by TrHSP70–13 (441 amino acids). Homology and phylogenetic studies showed that these sequences have the greatest homology to Takifugu bimaculatus. The mRNA of the two genes were constitutively expressed in all tested tissues of untreated T. rubripes, including brain, muscle, kidney, liver, spleen, gill, and intestine, with highest expression levels in the intestine. Three experimental groups were established in this study, the H1 group (20 °C), H2 group (24 °C) and H3 group (28 °C), in order to analyze the expressed levels of TrHSP70 in T. rubripes. To understand the role of HSPs in temperature stress in T. rubripes, we analyzed the expression levels of HSPs in T. rubripes at different temperatures. The results showed that the mRNA expression of the HSP gene were influenced by temperature stress. Gene expression of TrHSP70 and TrHSP60 were mainly up-regulated at 12 h and 48 h in the H1 group, respectively. Meanwhile, gene expression of TrHSP70 was highest at 48 h in the H3 group and TrHSP60 reached was lowest at 24 h in the H3 group. The results showed that the two genes were constitutively expressed genes, which play an important role in maintaining normal physiological function and stress responses. The results show that TrHSP70 and TrHSP60 are constitutively expressed genes, which plays important role in maintaining normal physiological function and coping with stress.http://www.sciencedirect.com/science/article/pii/S2352513421002817HSP60HSP70Takifugu rubripesMRNATemperature stresses
spellingShingle Kun-peng Fan
Ya-fang Liu
Zhi-qiang Zhang
Ying Liu
Peng-fei Liu
Molecular cloning, characterization and expression patterns of HSP70 and HSP60 in the pufferfish (Takifugu rubripes)
Aquaculture Reports
HSP60
HSP70
Takifugu rubripes
MRNA
Temperature stresses
title Molecular cloning, characterization and expression patterns of HSP70 and HSP60 in the pufferfish (Takifugu rubripes)
title_full Molecular cloning, characterization and expression patterns of HSP70 and HSP60 in the pufferfish (Takifugu rubripes)
title_fullStr Molecular cloning, characterization and expression patterns of HSP70 and HSP60 in the pufferfish (Takifugu rubripes)
title_full_unstemmed Molecular cloning, characterization and expression patterns of HSP70 and HSP60 in the pufferfish (Takifugu rubripes)
title_short Molecular cloning, characterization and expression patterns of HSP70 and HSP60 in the pufferfish (Takifugu rubripes)
title_sort molecular cloning characterization and expression patterns of hsp70 and hsp60 in the pufferfish takifugu rubripes
topic HSP60
HSP70
Takifugu rubripes
MRNA
Temperature stresses
url http://www.sciencedirect.com/science/article/pii/S2352513421002817
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