• 생명과학회지
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• 제16권5호
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• pp.729-733
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• 2006

크립토스포리디움의 활성 및 감염성 판정을 위해 Direct RT-PCR, 세포배양 후 RT-PCR 및 면역형광염색법을 비교한 결과는 다음과 같다. 1) 크립토스포리디움의 HSP70 gene에 대해 direct RT-PCR한 결과, 민감도가 매우 높아 저농도로 존재하는 환경시료에서의 크립토스포리디움 활성을 모니터링하는데 장점이 있을 것으로 보이나, 감염성의 판정은 알 수 없으며, 정량화가 안되는 단점이 있었다. 2) ${\beta}-tubulin$ gene에 대해 RT-nested PCR을 한 결과 크립 토스포리디움의 난포낭이 $1{\times}10^4$ 세포수정도 되어야 검출이 되는 것으로 나타나 HSP70 gene에 대한 RT-PCR결과와 비교할 때 10,000배 이상 민감도가 떨어지는 것으로 나타났다. 3) 세포배양 후 RT-PCR 또는 면역형광염색법을 이용할 경우에는 민감도가 direct RT-PCR보다 다소 떨어지는 단점이 있었으나 크립토스포리디움의 오염원이나 오염이 심한 지역의 감염성 조사에 적합할 것으로 나타났으나, 정량화가 필요한 경우에는 세포배양 후 면역형광염색법이 효과적일 것으로 나타났다.

• Animal cells and systems
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• 제14권4호
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• pp.267-274
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• 2010

The decrease in sperm quality under heat stress causes a great loss in animal husbandry production. In order to reveal the mechanism underlying the sperm quality decrease caused by heat stress, we first established a mild heat-treated mouse model. Then, the sperm quality was identified. Further, the testicular proteome profile was mapped and compared with the control using 2D electrophoresis and mass spectrometry. Finally, the differential expressed proteins involved in the heat stress response were identified by real-time PCR and Western blotting. The results showed that heat stress caused a significant reduction in mouse sperm quality (P<0.05). Further, 52 protein spots on the 2D gel were found to differ between the heat-shocked tissues and the control. Of these spots, some repair proteins which might provide some explanation for the influence on sperm quality were found. We then focused on Bag-1, Hsp40, Hsp60 and Hsp70, which were found to be differently expressed after heat shock (P<0.05). Further analysis in this heat-shocked model suggests numerous potential mechanisms for heat shock-induced spermatogenic disorders.

• Journal of Animal Science and Technology
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• 제64권6호
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• pp.1046-1062
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• 2022

This study investigated the effects of L-glutamine (Gln) supplementation on growth performance, physiological traits, heat shock proteins (HSPs), and gene expression related to muscle and adipose tissue development in Hanwoo steers under heat stress (HS) conditions. Eight Hanwoo steers (initial body weight [BW] 570.7 ± 43.6 kg, months of age 22.3 ± 0.88) were randomly separated into two groups, control and treatment, and supplied with the concentration (1.5% of BW kg/day/head) and rice straw (1.5 kg/day/head). The treatment group were fed the Gln supplementation (0.5% of concentration, as-fed basis) once a day at 08:00 h. Blood samples for the assessment of haematological and biochemical parameters and the separation of peripheral blood mononuclear cells (PBMCs) were collected four times, at 0, 3, 6, and 10 weeks of the experiment. Feed intake was measured daily. BW to analyze growth performance and hair follicle collection to analyze the expression of HSPs were executed four times at 0, 3, 6, and 10 weeks. To analyze gene expression, longissimus dorsi muscle samples were collected by biopsy at the end of the study. As a result, growing performance, including final BW, average daily gain, and gain-to-feed ratio, were not different between the two groups. Leukocytes including lymphocytes and granulocytes, tended to increase in the Gln supplementation group (p = 0.058). There were also no differences in biochemical parameters shown between the two groups, except total protein and albumin, both of which were lower in the Gln supplementation group (p < 0.05). Gene expressions related to muscle and adipose tissue development were not different between the two groups. As temperature-humidity index (THI) increased, HSP70 and HSP90 expression in the hair follicle showed a high correlation. HSP90 in the hair follicle was decreased in the treatment group compared with the control group at 10 weeks (p < 0.05). Collectively, dietary Gln supplementation (0.5% of concentration, as-fed basis) may not be influential enough to affect growth performance and gene expression related to muscle and adipose tissue development in steers. However, Gln supplementation increased the number of immune cells and decreased HSP90 in the hair follicle implying HS reduction in the corresponding group.

• Journal of Microbiology
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• 제34권2호
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• pp.158-165
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• 1996

The signal transduction pathways through the RAS gene product and adenyl cyclease play a critical role in regulation of the cell cycle in yeast, Saccharomyces cerevisiae. We examined the genetic relationship between the spt3 gene and ras/cAMP pathway. A mutation in the SPT3 gene suppressed cell cycle arrest at the G1 phase caused by either an inactivation of the RAS or CYR1 gene which encodes a yeast homologue of human ras proto-oncogene or adenyl cyclase, respectively. The phenotypes such as sporulation and heat shock resistancy, that resulted from a partial inactivation of the RAS or CYR1 genes, were also suppressed by the spt3 mutation. Expression of the SSA1 gene encoding one of th heat shock proteins (Hsp70) can be induced by heat shock or nitrogen starvation. Expression of this gene is derepressed in cry1-2 and spt3 mutants. The bcy 1 mutation repressed by the bcy1 mutation, but not in spt3 mutants. These results suggest that the SPT gene is involved in expression of genes that are affected by the RAS/cAMP pathway.

• 한국초지조사료학회지
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• 제33권3호
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• pp.159-166
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• 2013

We have previously investigated the proteome changes of rice leaves under heat stress (Lee et al. in Proteomics 2007a, 7:3369-3383), wherein a group of antioxidant proteins and heat shock proteins (HSPs) were found to be regulated differently. The present study focuses on the biochemical changes and gene expression profiles of heat shock protein and antioxidant genes in rice leaves in response to heat stress ($42^{\circ}C$) during a wide range of exposure times. The results show that hydrogen peroxide and proline contents increased significantly, suggesting an oxidative burst and osmotic imbalance under heat stress. The mRNA levels of chaperone 60, HSP70, HSP100, chloroplastic HSP26, and mitochondrial small HSP responded rapidly and showed maximum expression after 0.5 or 2 h under heat stress. Transcript levels of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and Cu-Zn superoxide dismutase (Cu-Zn SOD) showed a rapid and marked accumulation upon heat stress. While prolonged exposure to heat stress resulted in increased transcript levels of monodehydroascorbate reductase, peroxidase, glyoxalase 1, glutathione reductase, thioredoxin peroxidase, 2-Cysteine peroxiredoxin, and nucleoside diphosphate kinase 1, while the transcription of catalase was suppressed. Consistent with their changes in gene expression, the enzyme activities of APX and DHAR also increased significantly following exposure to heat stress. These results suggest that oxidative stress is usually caused by heat stress, and plants apply complex HSP- and antioxidant-mediated defense mechanisms to cope with heat stress.

• Preventive Nutrition and Food Science
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• 제17권4호
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• pp.274-279
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• 2012

For the quantitative analysis of genetically modified (GM) maize in processed foods, primer sets and probes based on the 35S promoter (p35S), nopaline synthase terminator (tNOS), p35S-hsp70 intron, and zSSIIb gene encoding starch synthase II for intrinsic control were designed. Polymerase chain reaction (PCR) products (80~101 bp) were specifically amplified and the primer sets targeting the smaller regions (80 or 81 bp) were more sensitive than those targeting the larger regions (94 or 101 bp). Particularly, the primer set 35F1-R1 for p35S targeting 81 bp of sequence was even more sensitive than that targeting 101 bp of sequence by a 3-log scale. The target DNA fragments were also specifically amplified from all GM labeled food samples except for one item we tested when 35F1-R1 primer set was applied. A reference plasmid pGMmaize (3 kb) including the smaller PCR products for p35S, tNOS, p35S-hsp70 intron, and the zSSIIb gene was constructed for real-time PCR (RT-PCR). The linearity of standard curves was confirmed by using diluents ranging from $2{\times}10^1{\sim}10^5$ copies of pGMmaize and the $R^2$ values ranged from 0.999~1.000. In the RT-PCR, the detection limit using the novel primer/probe sets was 5 pg of genomic DNA from MON810 line indicating that the primer sets targeting the smaller regions (80 or 81 bp) could be used for highly sensitive detection of foreign DNA fragments from GM maize in processed foods.

• 생태와환경
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• 제56권4호
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• pp.430-439
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• 2023

UV와 O₃는 정수처리 공정에 이용되는 물질로 유기물, 오염물질 및 미생물을 제거하는 것으로 많은 연구가 보고되어 있다. 본 연구에서는 저서성 대형무척추동물 중 오염지표 생물이며 실내 사육 중인 깔따구(C. flaviplumus, C. riparius)를 이용하여 UV와 O₃ 노출에 따른 생존율, 체 Fig. 5. Heatmap of relative mRNA expression of HSP70 in C. riparius and C. flaviplumus exposed to UV and O₃. 색 변화 및 열충격 단백질(Heat Shock Protein 70) 발현 변화를 관찰하였다. UV에 노출된 C. flaviplumus의 생존율은 24시간 후 70% 감소하였으며, C. riparius는 50%로 감소하였다. O₃에 노출된 C. flaviplumus는 생존율의 변화가 없었으며 C. riparius는 노출 10분 후 95%로 감소하였으나, 이후 노출 시간 동안 변화가 없었다. 또한, 체색 변화에서 두 종에 대한 UV와 O₃ 노출은 시간 의존적으로 붉은색의 체색이 옅어지는 현상을 관찰하였다. HSP70 유전자 발현에서 C. riparius는 UV 노출 후 대조군에 비해 발현이 증가하였으며, 노출 12시간 후에서 유의한 차이를 보였다(P<0.05). O₃에 노출된 C. flaviplumus는 대조군에 비해 상대적으로 낮은 발현이 관찰되었으며, 노출 10분과 1시간 후에서 유의한 차이를 보였다 (P<0.05). 이와 같은 결과는 UV와 O₃ 노출에 따른 깔따구에 대한 생태독성학적 영향을 보고하였다. 따라서 본 연구 결과는 정수장에서 이용되는 소독물질인 UV와 O₃가 정수장 내로 유입되는 깔따구에 주는 영향을 파악할 수 있는 기초자료로서 활용될 수 있을 것이다.

• 생태와환경
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• 제53권2호
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• pp.165-172
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• 2020

중금속은 다양한 경로를 통해 환경 중 배출되어 서식 생물에 노출되며 체내 다양한 생리학적 불균형을 유도한다. 본 연구에서는 수서생물지표종으로 이용되는 깔따구(Chironomus riparius)를 이용하여 야외 중금속(Al, Aluminum; Cr, Chromium; Cu, copper; Mn, Manganese; Zn, Zinc) 농도 노출에 따른 다양한 분자발현 반응과 상관성을 분석하였다. 생물 체내 분자 반응을 관찰하기 위해 heat shock protein 40, 70, 90 (HSP40, 70, 90), cytochrome 450(CYP450), Glutathione S-transferase (GST) and Serine-type endopeptidase (SP)를 이용하였다. 그 결과, 스트레스 분자마커로 이용되는 HSPs 유전자들은 중금속 노출된 개체들에서 대조군보다 높은 경향을 보였으며 Cu 노출 시 가장 높은 발현을 나타냈다. 해독에 관여하는 CYP450과 GST 유전자 발현 결과, Cr과 Cu에서는 다른 노출군에 비해 높은 발현 경향을 나타냈다. SP 유전자 발현 결과 Al을 제외한 모든 노출군이 대조군과 유사한 발현 패턴을 보였다. 이와 같은 연구 결과는 실내에서 환경 중 존재하는 실제 농도를 반영한 독성실험을 통해 노출물질과 농도에 따라 특이적으로 발현하는 분자마커 패턴을 보고하였다. 또한, 수생태계로 유입되는 중금속이 하천에 서식하는 생물에 주는 유해 영향에 대한 정보와 분자 지표 유전자들의 현장 적용 가능성을 보여준다.

• Journal of Oral Medicine and Pain
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• 제19권2호
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• pp.57-71
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• 1994

Gingival fibroblasts were cultured and subjected to the test of Northern blot analysis for the demonstration of various mRNA expression in response to the low level laser treatment. For duplication of in vivo. Wound healing process, fibroblasts were pretreated with proinflammatory cytokine interleukin-1$\beta$(IL-1$\beta$) or mitogenic substance phorbol 12-myristate 13-acetate(PMA) prior to laser irradiation. The results were as follows : 1. By the laser irradiation, the gene expression of collagen type I was markedly increased I n gingival fibroblasts, especially in the case of PMA pretreatment. The gene expression of collagen type IV, however, was not only affected by laser irradiation but also by chemical cell stimulation. 2. Oncogene v-myc expression was affected by both laser irradiation and IL-1$\beta$ or PMA stimulation, But v-fos gene expression was not detected in any case of this experimental system. 3. Heat shock gene(Hsp 70)was expressed constiutively, but slightly increased by laser irradiation. 4. mRNA of fibroblast growth factor(FGF) was induced by both laser irradiation and IL-1$\beta$ or PMA treatment.

• 한국가금학회지
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• 제47권2호
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• pp.95-105
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• 2020

본 연구는 한국토종닭 5품종 (재래황갈종, 토착로드종, 토착레그혼종, 토착코니시종 및 한국오계)에 대한 생산 특성 및 생리적 특성을 제시하고자 이들의 생산능력, 강건성 및 스트레스 반응 정도를 살펴보았다. 분석 대상으로 총 493수를 공시하고, 생산능력의 평가를 위해 생존율, 체중, 초산일령, 산란율 및 난중을 조사하였고, 강건성 및 스트레스 반응정도를 측정하기 위하여 텔로미어 함유율, heterophil-lymphocyte 비율, HSP-70, HSP-90α 및 HSP-90β 유전자 발현율을 분석하였다. 실험 결과, 생존율은 토착로드종이 99%로 가장 높았고, 한국오계가 91%로 가장 낮았다. 발생시부터 50주령까지의 체중은 토착코니시종, 토착로드종, 한국재래닭, 한국오계, 토착레그혼종의 순으로 변동없이 높게 나타났다. 22주령부터 50주령까지의 평균일계산란율은 토착레그혼종이 75%로 가장 높았고, 토착코니시종이 10%로 가장 낮게 나타났다. 생리적 지표 분석 결과, 텔로미어 함유율은 토착로드종이 가장 높았으며, 토착코니시종이 가장 낮았다. Heterophil-lymphocyte 비율 및 HSP-90β 유전자 발현율은 공히 토착코니시종이 가장 높았으며, 토착로드종이 가장 낮게 나타났다. 이러한 결과들은 토착로드종이 스트레스에 저항성이 높고, 토착코니시종이 스트레스에 가장 민감하게 반응함을 의미한다. 이상의 결과들을 종합할 때, 토착코니시종은 한국형 토종 육용 품종으로 개량함이 바람직하고, 토착레그혼종은 한국형 토종 난용 품종으로 개량하는 것이 바람직할 것으로 사료된다. 더불어 토착로드종은 양호한 생산능력과 뛰어난 강건성을 기반으로 한국형 토종닭 종계 모계통으로 활용하는 것이 바람직하리라 생각되고, 재래황갈종은 우수한 육질을 바탕으로 차별화 된 고급육질형 토종닭 품종으로 개량하는 것이 바람직하리라 사료된다.