• 한국가축번식학회지
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• 제4권1호
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• pp.20-27
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• 1980

This experiment was to study semen properties of Charolais for the a, pp.ication ot artificial insemination. The result obtained were summarized as follows: 1. In the preservation of liquid semen for 6 days, the survival rates of Charolais semen averaged 57.14% in skim milk solution and 58.17% in tris buffer solution. There were not differences. 2. Recovery of semen after thawing was vigorous in the semen that was diluted and frozen in 48 hrs. 3. The real rates of survival sperm for Charolais averaged 83% after living sperm was diluted and stained for 6 days. 4. Methylene blue reduction test diluted semen was fresh when it was diluted within 48 hrs. 5. If the diluted semen was preserved below 5$^{\circ}C$ in Charolais, the pH decreased by 0.2 in a day. 6. Diluted semen was more resistant to the cold shock than fresh semen. 7. In resistance against hot shock, sperm was almost dead in 20 minutes in 46.5$^{\circ}C$ in diluted semen, while it was dead in 30 minutes in 42.5$^{\circ}C$ in diluted semen. 8. In examination of morphological changes of sperm acrosome for 6 days, normal sperm in skim milk solution and tris buffer solution was 80% and 76.97% respectively, swelling sperm 12.8% and 15.27%, deficient sperm 0.6% and 0.97% abnormal staining 3.07% and 5.25%, immature sperm 0.28%, and 0.23%, whereas other abnormal sperm was 1.28% and 1.42%.

• KSBB Journal
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• 제29권1호
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• pp.22-28
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• 2014

In this study, we investigate the effect of pretreatment method on lipid extraction from Enteromorpha intestinalis using physical, thermo-chemical, and enzymatic process such as ultrasonication, high temperature treatment, freezing, microwave irradiation, osmotic shock, pH shock, homogenizing, and enzymatic treatment. In pretreatment with separated lipid extraction, the high extraction yield was obtained by high temperature treatment ($121^{\circ}C$ for 5 min) with 0.1 N HCl, which is 1.4 times higher than that of control. In pretreatment with direct lipid extraction, the high extraction yields were obtained by 0.1 N HCl pretreatment, microwave irradiation (700W, 1 min with twice), and 10% NaCl pretreatment, which is 1.45 times higher than that of control. In the result of enzymatic pretreatment with 17 kinds of enzymes, Cellic CTec II showed the high extraction yield of 5.3%, and which is 1.9 times higher than that of control. Moreover, the extraction yield was increased by the increase of enzyme amounts. In 10% enzyme amount, about 5.8% yield was obtained.

• 미생물학회지
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• 제26권4호
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• pp.291-297
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• 1988

신호배열 돌연변이인 rbsB 103는 전구체 리보스 결합단백질을 세포질내에 축적시키고, rbsB 106 복귀유전자는 이 전구체를 숙성 가능하게 하여 페리플라슴으로 수송되게 한다.(Iida el at., 1985, Park, el at., 1988). 본고에서는 rbsB 유전자의 세 allele, rbsB, rbsB 103와 rbsB 106를 이들이 코딩하는 단백질을 대량생산 하그l자 람다 P 프로모터 조절하에 클론하고 나아가서 다섯종의 단백질을 순수정제하였다.. 전구체 단백질의 pI는 8.0, 숙성단백질의 $P_{L}$는 7.5임을 밝혔다. 순수정제된 단백질의 아마노 말단의 아미노산 배열을 결정하여 DNA 염기서열로부터 밝혀진 아미노산 변화를 확인하였다.

• 한국식품과학회지
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• 제40권6호
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• pp.712-716
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• 2008

Salmonella로부터 식품 안전성을 높이기 위한 보존법의 병용처리에 의한 효과를 평가하고자 S. Typhimurium을 열과 산, 산화제 등으로 연속 처리한 후 생균수를 측정하여 효과를 분석하였다. 그리고 열충격에 의하여 S. Typhimurium 내에 발현되거나 억제되는 단백질을 이차원 전기영동과 MALDI-TOF 질량분석기로 분석하였다. 열처리된 S. Typhimurium은 초산과 염산의 pH 4에서의 생균수가 1.3-1.8 log CFU/mL가 줄었고 비열처리 S. Typhimurium은 생균수가 약 5 log CFU/mL가 감소하였다. 열처리 S. Typhimurium은 butyl hydrogen peroxide와 과산화수소에서 생균수가 1.1-1.7 log CFU/mL가 줄었으나 비열처리 S. Typhimurium은 5.4-5.6 log CFU/mL 감소하였다. 충분하지 않은 사멸 열처리는 S. Typhimurium의 생존력을 증가시키고 산과 산화제 등의 보존제에서 저항성이 커지는 것을 알 수가 있었다. 이차원 전기영동과 MALDI-TOF 질량분석에 의한 발현 단백질 분석 결과 비열처리 S. Typhimurium은 17개의 단백질이 검출되었고 열처리 S. Typhimurium에는 13개의 단백질만 검출되었다. 이들 중에 열충격 단백질로 알려진 DnaK, small heat shock protein 등이 검출되었고 이들이 산과 산화제에서의 생존 저항성 증가와 관련이 있을 것으로 보인다. 그러므로 열처리를 포함하는 hurdle technology를 적용하여 식품을 보존처리할 때 다른 보존제에 대한 교차보호성이 증가되는 사실을 고려하여 적절한 열처리가 고려되어야 된다는 것을 알 수 있었다.

• Clinical and Experimental Reproductive Medicine
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• 제26권2호
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• pp.225-230
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• 1999

In mouse, the heat shock protein 70-2 (hsp70-2) is found to have special function in spermatogenesis. Based on the observation, the hypothesis that human hspA2 (human gene; 98.2% amino acid homology with hsp70-2) might have important function in spermatogenesis in human testes was proposed. To test the hypothesis, we examined the expression of hspA2 in human tissues. Expression vector pDMC4 for expression of the human hspA2 protein using pTricHisB (invitrogen, USA) was constructed and the expressed hspA2 protein was cross-reacted with antiserum 2A raised against mouse hsp70-2 protein. Based on the cross-reactivity, we determined the expression level of hspA2 protein in human tissues by western blot analysis using the antiserum 2A. We demonstrated that antiserum 2A antibodies detected human hspA2 protein with specificity which was produced in the E.coli expression system. On Western blot analyses, significant hspA2 expression was observed in testes with normal spermatogenesis, whereas a low level of hspA2 was expressed in testis with Sertoli-cell only syndrome. Also, a small amount of hspA2 was detected in breast, stomach, prostate, colon, liver, ovary, and epididymis. These results demonstrate that the hspA2 protein is highly expressed in male specific germ cells, which in turn suggests that hspA2 protein might playa specific role during meiosis in human testes as suggested in the murine model. However, further studies should be attempted to determine the function of hspA2 protein in human spermatogenesis.

• 한국동물학회지
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• 제26권2호
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• pp.69-81
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• 1983

溫熱處理가 SCK 腫瘍細胞에 미치는 生物學的 環境을 몇가지 측면에서 檢討하였다. 첫째로, 溫熱處理가 in vitro와 in vivo에서 어떤 차이가 있는 지를 알아보기 위해서 生殘曲線을 구하여 비교하여 본 바, in vivo의 경우에 더욱 큰 腫瘍細胞 致死效果를 보았다. 둘째로, 이러한 細胞致死作用의 차이가 나타나는 原因을 알아보기 위해서 溫熱處理가 腫瘍組織과 筋肉組織의 溫度와 pH에 어떤 차이를 나타내는 지를 측정하여 본바, 腫瘍의 경우 溫度는 筋肉보다 높아졌으며, pH는 초기의 上昇을 제외하고는 계속해서 낮은 狀態를 유지했다. 이와 같은 腫瘍내의 酸性狀態가 溫熱處理의 細胞致死效果를 增强시키는 것으로 보였다. 셋째로, 이와같은 腫瘍내 pH의 酸性化의 原因을 규명해 보기 위해서 腫瘍과 肝 속의 乳酸含量을 정량하여 본 바, 腫瘍의 경우 溫熱處理 후 1시간에는 增加하였으나 그 후에는 계속 減少된 상태를 유지했다. 이에 반해서 肝의 경우 5시간에 最大値를 보였고 24시간 동안 계속해서 上昇된 값을 유지했다. 이러한 腫瘍내 乳酸含量의 減少는 예기치 못했던 결과였는데, 이는 溫熱處理로 말미암아 腫瘍내의 細胞가 계속해서 死滅되는 결과로 腫瘍 단위 重量당 乳酸量의 감소를 가져오는 것으로 풀이할 수 있었다. 넷째로, 溫熱處理에 의한 細胞致死效果의 표적을 찾아 보기 위한 한가지 試圖로서 膜表面蛋白質의 變化를 관찰하였던 바, 高分子蛋白質의 減少와 더불어 低分子蛋白質의 增加라는 一般的인 現象을 볼 수 있었고, 특히 70K 단백질이 特異的 增加를 보인 것과 이의 증가가 $41^\\circC$에서 最大의 값을 보인 것은 興味로운 일이었다. 溫熱處理가 腫瘍내 pH의 酸性化를 招來하는 原因과 膜表面蛋白質에 特異한 影響을 미치는 機作에 대해서는 앞으로 더욱 많은 硏究가 傾注되어야 할 興味있는 問題로 생각된다.

• Bulletin of the Korean Chemical Society
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• 제33권8호
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• pp.2508-2512
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• 2012

Cold shock proteins (CSPs) are a family of proteins induced at low temperatures. CSPs bind to single-stranded nucleic acids through the ribonucleoprotein 1 and 2 (RNP 1 and 2) binding motifs. CSPs play an essential role in cold adaptation by regulating transcription and translation via molecular chaperones. The solution nuclear magnetic resonance (NMR) or X-ray crystal structures of several CSPs from various microorganisms have been determined, but structural characteristics of psychrophilic CSPs have not been studied. Therefore, we optimized the purification process to obtain highly pure Lm-Csp and determined the three-dimensional structure model of Lm-Csp by comparative homology modeling using MODELLER on the basis of the solution NMR structure of Bs-CspB. Lm-Csp consists of a ${\beta}$-barrel structure, which includes antiparallel ${\beta}$ strands (G4-N10, F15-I18, V26-H29, A46-D50, and P58-Q64). The template protein, Bs-CspB, shares a similar ${\beta}$ sheet structure and an identical chain fold to Lm-Csp. However, the sheets in Lm-Csp were much shorter than those of Bs-CspB. The Lm-Csp side chains, E2 and R20 form a salt bridge, thus, stabilizing the Lm-Csp structure. To evaluate the contribution of this ionic interaction as well as that of the hydrophobic patch on protein stability, we investigated the secondary structures of wild type and mutant protein (W8, F15, and R20) of Lm-Csp using circular dichroism (CD) spectroscopy. The results showed that solvent-exposed aromatic side chains as well as residues participating in ionic interactions are very important for structural stability. Further studies on the three-dimensional structure and dynamics of Lm-Csp using NMR spectroscopy are required.

• 미생물학회지
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• 제34권1_2호
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• pp.43-50
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• 1998

여러가지 환경 stress에서 유도되는 폐렴구균 열충격단백질 GroEL의 특성에 대하여 검토하였다. 비병원성인 폐렴구균(Streptococcusa pneumoniae)을 사용하여 stress 조건을 설정하고 이 설정된 조건에서 stress에 의해 유도되는 단백질을 $^{35}S$]-methionine으로 표지하여 autoradiography를 실시하였다. 열충격을 가했을 때 유도되는 단백질(65, 73, 84-kDa 등) 중 65 kDa의 단백질(hsp65)을 DEAE-Sepharose ion exchange 및 ATP-agarose affinity chromatography를 이용하여 분리 정제하였으며 hsp65에 대해 생성된 항체를 이용하여 immunoblot을 실시하였을 때 약 60 kDa의 대장균 단백질과 반응하였으며 정제된 폐렴구균 hsp65가 대장균의 anti-GroEL monoclonal 항체와 반응함으로써 폐렴구균 hsp65가 대장균GroEL과 유사한 단백질임을 확인하였다.

• Journal of Plant Biotechnology
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• 제44권3호
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• pp.287-295
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• 2017

Water temperature is one of the major factors that impacts the growth and life cycle of Pyropia tenera, one of the most valuable and cultivated marine red algae belonging to Bangiales (Rhodophytes). We analyzed transcriptome from gametophyte of P. tenera under normal and high temperature conditions, and identified four small heat shock proteins (sHSPs). They have no significant amino acid sequence homology with known proteins in public databases except PhsHSP22 from Pyropia haitanensis. PtsHSP19.3 gene responded to high temperature but slightly or not to desiccation, freezing or high salt condition. When the PtsHSP19.3 gene was overexpressed in Chlamydomonas reinhardtii, transformed Chlamydomonas lines revealed much higher growth rate than that of control cells under heat stress condition. Transformed cells also grew well in those of the control cell onto the medium containing high salt or $H_2O_2$. When the PtsHSP19.3 was fused to GFP and introduced into tobacco protoplast, fluorescence was detected at several spots. Results indicate that PtsHSP19.3 may form super-molecular assembles and be involved in tolerance to heat stress.

• 환경위생공학
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• 제14권1호
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• pp.54-61
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• 1999

The industrial techniques of ultrasound have been used in the various fields, such as cleaning, medical surgery, emulsification, cell disruption etc. Especially the application of cell disruption was interested in the field of disinfection process in water by ultrasonic irradiation. It has been recognized that the ultrasounds are irradiated in aqueous solution, cavitation bubbles are generated and shock waves of high temperature and pressure are emitted as the bubbles are developed and finally broken, which function as a energy source to promote reaction efficiencies of various kinds of chemical reactions such as disinfection reaction in water. Therefore, this study was performed to apply the ultrasound for the disinfection method of infected drinking raw water and to discuss the limiting factors such as pH, sample volume and reaction temperature influenced on the removal efficiency of E. coli from experimental analysis of the results obtained in bench-scale plant. For the experiments to measure the influence of reaction parameters in the ultrasonic disinfection process, escalated reactivity of aqueous solutions was excellent when pH in aqueous solution was low, and sample volume was small. On the contrary, the reactivity of disinfection became elevated when reaction temperature was high. It was found that the rate constant of disinfection reaction was applied by Chick's law, reaction kinetics of Chick's law was irreversible and pseudo-first order at all the tested conditions.As a conclusion it appeared that the technology using ultrasonic irradiation can be applied to the treatment of disinfection in infected water which are difficult to be treated by conventional methods.