• Korean Journal of Ichthyology
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• v.19 no.4
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• pp.266-273
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• 2007

Heat shock proteins (HSPs) are a family of highly conserved proteins playing an important role in the functioning of unstressed and stressed cells. The HSP70 family, the most widely studied of the hsps, is constitutively expressed (hsc70) in unstressed cells and is also induced in response to stressors (hsp70), especially those affecting the protein machinery. The HSP/HSC70 proteins act as molecular chaperones and are crucial for protein functioning, including folding, intracellular localization, regulation, secretion, and protein degradation. Here, we report the identification and characterization of the putative amino acid sequence deduced from one cDNA clone identified as heat shock protein 70. The alignment showed that the putative sequence is 100% identical to the heat shock protein 70 cognate (HSC 70) of olive flounder. The 5'-flanking region sequence (approximately 1 kb) ahead of the hsc70 gene was cloned by genome walking and a putative core promoter region and transcription elements were identified. We characterized the promoter of the olive flounder hsc70 gene by examining the ability of 5'-upstream fragments to drive expression of green fluorescent protein (GFP) in live embryos.

• International Journal of Industrial Entomology and Biomaterials
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• v.29 no.2
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• pp.145-152
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• 2014

The variation in the level of immune response related gene expression in silkworm, Bombyx mori following infection with Bombyx mori nucleopolyhedrovirus (BmNPV) was analyzed at different time intervals. The occlusion bodies of BmNPV orally inoculated to the two most divergent silkworm races viz., Sarupat (resistant to BmNPV infection) and CSR2 (susceptible to BmNPV infection) were subjected to oral BmNPV inoculation. The expression profile of gp 41 gene of BmNPV in the Sarupat and CSR2 races revealed that the virus could invade the midguts of both susceptible and resistant races. However, its multiplication was significantly less in the midgut of resistant race, while, in the susceptible race, the viral multiplication reached maximum level within 12 h. These findings indicate that potential host genes are involved in the inhibition of viral multiplication within larval midgut. The immune response genes arylphorin, cathepsin B, gloverin, lebocin, serpin, Hsp 19.9, Hsp 20.1, Hsp 20.4, Hsp 20.8, Hsp 21.4, Hsp 23.7, Hsp 40, Hsp 70, Hsp90 revealed differential level of expression on NPV infection. The gloverin, serpin, Hsp 23.7 and Hsp 40 genes are significantly up-regulated in the resistant race after NPV infection. The early up-regulation of these genes suggests that these genes could play an important role in baculovirus resistance in the silkworm, B. mori.

• The Korean Journal of Zoology
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• v.39 no.2
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• pp.123-131
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• 1996

In this study, we examined the expression of heat shock proteins (HSPs) in fish cell line CHSE-2lnl and human HeLa cells exposed to heat shock. In fish CHSE-214 cells HSP70 was the major polvpeptide induced by an elevated temperature or an amino acid analog, while in HeLa cells HSP90 as well as HSP70 were prominently enhanced in response to these stresses. Pretreatment of actinomvcin D prior to heat shock completely inhibited the induction of fish HSP70, indicating the transcriptional regulation of fish HSP70 gene expression. In HeLa and CHSE-214 cells either recovering from heat shock or experiencing prolonged heat shock, attenuation in the HSP90 a'nd HSP70 induction occurred but both induction and repression of HSP70 synthesis appear 19 precede those of HSP90. Moreover, attenuation did not occur in the syntheses of 40 kDa and 42 kOto proteins which were only induced in CHSE-214 cells. The enhanced syntheses of these he proteins continued as long as CHSE-214 cells were Siven heat shock. These results suggest that down-regulation of HSP syntheses during prolonged heat shock may be controlled by several different. as vet undefined, mechanisms.

• Journal of Life Science
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• v.28 no.6
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• pp.639-647
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• 2018

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 $12^{\circ}C$) for sampled livers revealed that expression gradually increased at $12^{\circ}C$ compared to $21^{\circ}C$. 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.

• Journal of Sericultural and Entomological Science
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• v.39 no.1
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• pp.36-43
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• 1997

In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.4
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• pp.515-523
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• 2020

Basal and heat shock-induced mRNA expression patterns of major heat shock protein (HSP) genes, including those encoding heat shock protein (HSP) 90, HSP70, HSP70-12A, heat shock inducible protein 70 (HSIP70), heat shock binding protein 1 (HSPBP1), HSP60, and HSP40 were examined in the gill and hepatopancreas of 1-year-old diploid and triploid abalone Haliotis discus hannai juveniles. Under non-stimulated conditions at 19℃, triploid abalones displayed, in general, higher mRNA levels of various HSPs (HSP70, HSIP70, HSPBP1, HSP70-12A, and HSP60 in the gill and HSIP70, HSPBP1, and HSP60 in the hepatopancreas) than did communally cultured diploids. Conversely, only the hepatopancreatic expression of HSP70-12A was higher in diploids than in triploids. However, the fold changes in gene expression in response to an acute thermal challenge (elevation from 19 to 30℃) were generally greater in diploids than in triploids, such that the difference in basal expression was diminished, weakened, or even reversed after heat shock treatment. However, unlike other HSP genes, the basal expression of HSP60 (higher in 3N) was more pronounced after heat shock treatment. Collectively, the results of this study suggest that triploid abalones have different capacities for not only basal expression but also the heat-induced expression of HSPs in an HSP member-dependent manner.

• Parasites, Hosts and Diseases
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• v.55 no.4
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• pp.367-374
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• 2017

Despite the broad distribution of leishmaniasis among Iranians and animals across the country, little is known about the genetic characteristics of the causative agents. Applying both HSP70 PCR-RFLP and sequence analyses, this study aimed to evaluate the genetic diversity and phylogenetic relationships among Leishmania spp. isolated from Iranian endemic foci and available reference strains. A total of 36 Leishmania isolates from almost all districts across the country were genetically analyzed for the HSP70 gene using both PCR-RFLP and sequence analysis. The original HSP70 gene sequences were aligned along with homologous Leishmania sequences retrieved from NCBI, and subjected to the phylogenetic analysis. Basic parameters of genetic diversity were also estimated. The HSP70 PCR-RFLP presented 3 different electrophoretic patterns, with no further intraspecific variation, corresponding to 3 Leishmania species available in the country, L. tropica, L. major, and L. infantum. Phylogenetic analyses presented 5 major clades, corresponding to 5 species complexes. Iranian lineages, including L. major, L. tropica, and L. infantum, were distributed among 3 complexes L. major, L. tropica, and L. donovani. However, within the L. major and L. donovani species complexes, the HSP70 phylogeny was not able to distinguish clearly between the L. major and L. turanica isolates, and between the L. infantum, L. donovani, and L. chagasi isolates, respectively. Our results indicated that both HSP70 PCR-RFLP and sequence analyses are medically applicable tools for identification of Leishmania species in Iranian patients. However, the reduced genetic diversity of the target gene makes it inevitable that its phylogeny only resolves the major groups, namely, the species complexes.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.2
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• pp.280-285
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• 2006

The current study was conducted to investigate the relationship between stress related gene and meat quality in pigs. A total number of 212 three-way cross bred (Landrace-$Yorkshire{\times}Duroc$) and 38 Duroc were sampled from the Korean pig industry to determine genotype frequency of porcine stress syndrome (PSS) and heat shock protein 70 kDa (HSP70) genes and their relationship with carcass traits and longissimus meat quality. Screen of HSP70 was performed by the single strand conformation polymorphism (SSCP) technique. Based on the analysis of restriction fragment length polymorphism (RFLP) in ryanodine receptor 1 (RYR1) gene, genetic disorder of PSS was related to a mutation at $18,168^{th}$ (C to T) of exon 17. There was no significant difference in ultimate meat pH and backfat thickness between HSP70 K1-AA type and -BB type in pure Duroc breed. In Landrace-$Yorkshire{\times}Duroc$ (L-$Y{\times}D$) cross bred pig, our results indicated that HSP70 derivate type in Duroc had a limited effect on backfat thickness, but L-$Y{\times}D$ type had a noticeable linkage with HSP70 K1-AA and K3-AB. This tendency was also observed in hot carcass weight where HSP70 K1-AA and K3-AB resulted in heavier weight with 86.3 kg compared to HSP70 K1-AB and K3-BB of 74.3 kg. Results imply that stress related HSP70 genotype has a potential association with backfat thickness and carcass weight.

• Korean Journal of Environmental Biology
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• v.33 no.4
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• pp.450-458
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• 2015

The heat shock proteins (Hsps), one of the most highly conserved groups of proteins, play crucial roles in protecting cells against environmental stressors, such as temperature, salinity, heavy metals and pathogenic bacteria. The glutathione S-transferases (GST) have important role in detoxification of oxidative damage, environmental chemicals and environmental stress. The purpose of this study is to investigate the gene expression of Hsp70 and GST on change of temperature and salinity in Mytilus coruscus. The M. coruscus was cultured in incubator of separate temperature and salinity (8, 20, $30^{\circ}C{\times}20$‰, 25‰, 30‰) for 28 days. Ten individuals in each group were selected after each 14 and 28 days exposure. Results that the expression of Hsp70 mRNA was no significant changed in M. coruscus exposed to temperature ($8^{\circ}C$, $20^{\circ}C$, $30^{\circ}C$) and salinity (20‰, 25‰, 30‰) for 14 days. Whereas the expression of Hsp70 mRNA was increased in exposure to temperature $30^{\circ}C$ and salinity (20‰, 25‰, 30‰) for 28 days. The expression of GST mRNA was increased in exposure to temperature $30^{\circ}C$, salinity (25‰, 30‰) for 14 days and temperature ($8^{\circ}C$, $20^{\circ}C$, $30^{\circ}C$), salinity (20‰, 25‰, 30‰) for 28 days. These results suggest that Hsp70 and GST were played roles in biomarker gene on the thermal and salinity stress.

• Animal cells and systems
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• v.14 no.1
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• pp.17-23
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• 2010

We isolated warm temperature acclimation-related protein 65-kDa (Wap65) cDNA from the liver of olive flounder and investigated the mRNA expression of Wap65 and HSP70 in olive flounder exposed to osmotic (17.5, 8.75, and 4 psu) and thermal stress (25 and $30^{\circ}C$). The mRNA expression of Wap65 and HSP70 was increased by thermal stress. The mRNA expression of HSP70 was also increased by osmotic stress, whereas no significant change in Wap65 expression was detected. These results indicate that Wap65 mRNA expression occurs specifically in response to increases in water temperature, but not in response to osmotic stress. Plasma cortisol levels were also increased by osmotic and thermal stress. We also utilized the stress hormone cortisol to examine whether Wap65 expression is thermal-stress-specific. Cortisol treatment increased HSP70 mRNA expression in vitro, but had no significant effect on Wap65 mRNA expression. Thus, thermal stress, but not osmotic stress, induces Wap65 expression.