Heat shock proteins (HSPs) are highly conserved cellular proteins that contribute to adaptive responses of organisms to a variety of stressors. In response to stressors, cellular levels of HSPs are increased and play critical roles in protein stability, folding and molecular trafficking. The mRNA expression pattern of two well-known heat shock protein transcripts, HSP70 and HSP90 were studied in two tissues of nerve ganglia, cerebral ganglion and pleuropedal ganglion of Pacific abalone (Haliotis discus hannai). It was observed that both HSP70 and HSP90 transcripts were upregulated under heat stress in both ganglion tissues. Expression level of HSP70 was found higher than HSP90 in both ganglia whereas cerebral ganglion showed higher expression than pleuropedal ganglion. The HSP70 and HSP90 showed higher expression at Day-1 after exposed to heat stress, later decreased at Day-3 and Day-7 onwards. The present result suggested that HSP70 and HSP90 synthesize in nerve ganglion tissues and may provide efficient protection from stress.
Glutathione S-transferase (GST) is a key gene involved in multiple stress tolerance in all living organisms, though it is still to be disclosed the gene function in teff grass [Eragrostis tef (Zucc.)Trotter].The objectives of this study were to clone and molecular characterization of GST gene in teff grass. We characterized GST1 from teff grass (EtGST1), it composed of a 645-bp open reading frame (ORF) that encoded 195 amino acid residue. Further, we transformed EtGST1 in E.coli BL21 (DE3) cells. This recombinant EtGST1 in E.coli BL21(DE3) induced at 37°C temperature. In addition, Growth of cells overexpressing EtGST1 rapidly increased in the presence of polyethylene glycol (5%), heat (46°C), NaCl (0.6%), and arsenic (1 mM) than that of cells harboring an empty vector. These results suggest that EtGST1 would be suitable candidate for improving tolerance in forages and/or grasses species against multiple abiotic stresses.
Arabidopsis nucleoside diphosphate kinase 2 (AtNDPK2) is an upstream signaling molecule that has been shown to induce stress tolerance in plants. In this study, the AtNDPK2 gene, under the control of a stress-inducible SWPA2 promoter, was introduced into the genome of tall fescue (Festuca arundinacea Schreb.) plants. The induction of the transgene expression mediated by methyl viologen (MV) and NaCl treatments were confirmed by RT-PCR and northern blot analysis, respectively. Under salt stress treatment, the transgenic tall fescue plants (SN) exhibited lower level of H2O2 and lipid peroxidation accumulations than the non-transgenic (NT) plants. The transgenic tall fescue plants also showed higher level of NDPK enzyme activity compared to NT plants. The SN plants were survived at 300 mM NaCl treatment, whereas the NT plants were severely affected. These results indicate that stress-inducible overexpression of AtNDPK2 might efficiently confer the salt stress tolerance in tall fescue plants.
본 연구는 열 스트레스 하에서 오리사료 내 대사에너지(ME) 수준이 오리의 간, 십이지장 융 모, 미생물, 유전자 조절에 미치는 영향을 조사하였다. 총 240마리의 육용 오리 채리밸리(Anas platyrhynchos)를 4처리구로 완전임의배치 한 후 42일 동안 사육하였다. 처리구는 ME 2900 kcal/kg, ME 3000 kcal/kg, ME 3100 kcal/kg 및 ME 3200 kcal/kg로 구분하였다. 간 조직은 처리구 사이의 차이가 없었고, 십이지장 융모 및 창자샘 길이는 ME 3000과 비교할 때 2900은 10.58% 감소하였으나 3100, 3200과의 사이에 차이는 없었다. 맹장 Latobacillus는 ME 3000과 비교할 때 2900은 9.47% 감 소하였으나 3100, 3200은 각각 2.52, 3.24% 증가하였다. Total aerobic bacteria, E. coli, Coliform bacteria는 ME 3000과 비교할 때 2900은 증가하였으나 3100, 3200은 차이가 나타나지 않았다. 간에서 HSP (heat shock proteins)-mRNA 중 HSP 90-α는 ME 3000과 비교할 때 2900은 48.60% 감소하였 으며 3100, 3200은 차이가 없거나 증가하였다.
The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms.
In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions.
Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.
The sweetpotato whitelfy Bemisia tabaci distribute worldwide and infests more than 500 species of plants. To determine nutritional stress of whiteflies at molecular level, we identified a full cDNA of glucose regulated protein 78 (grp78) which is known to be respond to nutritional restriction in vertebrate species. GRP78 of B. tabaci was highly conserved motifs of the HSP70 family and the C-terminal motif of KDEL characteristic of endoplasmic reticulum-specific HSPs. Real-time RT-PCR analysis showed that the grp78 level was not changed by thermal stress treatment from 4°C to 40°C for 1 h. However, the grp78 level was proportionally increased to the ingestion of a sucrose solution ranging in concentrations from 0% to 30% in a Parafilm feeding chamber. In addition, the grp78 levels were various by the ingestion of leaves of 10 different plants for 24 h; its level was higher with eggplant and pepper but lower with rice and apple. Our study suggests that the grp78 may have a role for cellular chaperones in relation to nutritional uptake of B. tabaci.
저온 스트레스에 대한 내성을 지닌 신품종 톨 페스큐를 개발할 목적으로 CaMV35S 프로모터 하류에 NDP kinase 2 유전자가 항상적으로 발현하도록 제작한 벡터를 Agrobacterium법을 이용하여 톨 페스큐에 도입하였다. Hygromycin이 첨가된 선발배지에서 내성을 나타내며 재분화된 형질전환 식물체를 pot로 이식하여 기내순화 시킨 후, Southern blot 분석을 실시하여 본 결과, NDP kinase 2 유전자가 형질전환 식물체의 g
Environmental stress is a growing problem for the productivity of forage crops. Reactive oxygen species (ROS) formation due to several abiotic stresses is a fundamental process that interrupts several physiological processes and causes a significant reduction on growth and yield of many forage crops. Molecular breeding such as genetic transformation has become a popular biotechnological tool for improving forage quality as well as improves tolerance to various abiotic stresses. As a first step of genetic transformation in tall fescue, we established an efficient Agrobacterium-mediated genetic transformation protocol using mature seed-derived embryogenic callus. After optimization of the transformation system, several genes of interest have been used to generate abiotic stress tolerant forage grasses. We generated transgenic tall fescue plants expressing NDPK genes under the control of the oxidative stress-inducible SWPA2 promoter. Transgenic plants showed enhanced tolerance to several abiotic stresses. Results in the current study, suggest that NDPK mediated multiple stress tolerance by increasing the expression of genes involved in antioxidant and protective functions, possibly through activation of an MAPK cascade.
This study was performed to investigate the relationship between PSS-HSP70 gene polymorphism and artificial insemination (AI) reproductivity in the pigs. The RFLP polymorphism of PSS and the SSCP polymorphisms of HSP70 K1, K3 and K4 PCR product were detected different patterns. In the experiment for AI of fresh semen, spring and fall season showed higher litter size born of 10.89 head than 10.47 head of summer season. Landrace was showed higher litter size of 9.96 head than that of Duroc and Yorkshire (p<0.05). Stress relating PSS and HSP70 polymorphism of PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showd a highest litter size born of 10.97 head and litter size born alive of 10.69 head than that of the other polymorphisms(p<0.05). In the experiment for AI of frozen semen, effects of season and pig breeds were not showed for litter size born. The stress relating polymorphism of PSS-Carrier, HSP70 K1-BB, K3-BB, K4-AB showed highest litter size born of 11.29 head and litter size born alive of 10.82 head and PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showed the lowest litter size born of 8.48 head and litter size born alive of 7.33 head than that of the other polymorphisms(p<0.05). These results suggest that AI litter size born for the stress of forzen thawed semen may be affected by PSS and HSP70 polymorphism in pigs.