• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
• /
• pp.5-5
• /
• 2002

Nucleoside diphosphate kinases (NDPKs) are conserved through evolution and have been shown to be involved in various biological phenomena. By functional screening in yeast, we identified a new member of the NDPK family, ㎚23-M5, which encodes a 211-amino acid protein with 86% identify to the human homolog, ㎚23-H5. Northern blot analysis reveals that ㎚23-M5 encodes two transcripts of 0.8 and 0.7 kb, which are highly and specifically expressed in adult testis. Reverse transcriptase polymerase chain reaction analysis shows that nm23-M5 first appears in pachytene spermatocytes and increase chain reaction in abundance through subsequent stages. (omitted)

• Ocean and Polar Research
• /
• 제23권4호
• /
• pp.337-345
• /
• 2001

A Cd-binding protein was identified in the renal cytosol of the Antarctic clam Laternula elliptica which naturally contains high concentrations of Cd. The Cd-binding protein showed similar characteristics of metallothionein (MT) in molecular weight (about 10-12 kDa) and low spectral absorbance at 280 nm with relatively high absorbance at 254nm. Results of immuno-histochemical staining suggested that the MT-like Cd-binding protein was mainly located in the epithelial cells of the kidney. The MT-like protein was a major ligand of cytosolic Cd as shown in the elution profiles of chromatography and may play an important role in Cd sequestration and accumulation in L. elliptica kidney. A considerable amount of Cd was also found to be associated with particulate fraction, indicating the sequestration to particulate fraction is as important as binding to the cytosolic MT-like protein in Cd accumulation in the kidney.

• Bulletin of the Korean Chemical Society
• /
• 제23권12호
• /
• pp.1724-1728
• /
• 2002

The adsorption of proteins on the surface of glass slides is essential for the construction of protein chips. Here, we report that a Histidine (His)-tagged protein protein has been efficiently adsorbed on glass coated with nickel. A variety of nickel chloride-coated plates were prepared by the spin-coating method and adsorbed to the His-tagged protein. When the protein was adsorbed onto the surface of a variety of nickel chloride-coated glass slides, the efficiency of protein adsorption was dependent upon the coating conditions such as nickel chloride concentration, the spin speed and the drying temperature. The slides appropriate for protein adsorption were obtained when the slides were coated with 11%(w/w) of $NiCl_2$ at the spin speed of 4000 rpm for 20 sec and then dried at higher than 40°C. The physical properties of their nickel chloride thin layer were characterized by scanning electron microscopy. x-ray diffraction and atomic force microscopy, finding that the nickel chloride particles were around 10 nm in diameter and uniformly crystallized at 101 faces. These results show that nickel chloride-coated slides prepared by the spin-coating method are utilizable for the construction of Histagged protein chips.

• 분석과학
• /
• 제24권3호
• /
• pp.193-199
• /
• 2011

본 실험은 AP Tech사에서 생산된 rProtein A의 단기 안정성실험을 통하여 저장방법 및 사용기간 설정과 경시변화에 따른 품질의 안정성을 평가를 목적으로 한다. 즉, 생산된 rProtein A의 보관조건인 $-20^{\circ}C$에서 냉동시킨 시료를 사용 혹은 실험 후 냉장보관($4^{\circ}C$) 했을 때 얼마의 시간동안 안정성을 확보하는가를 보자고 하였으며, 실험기간은 초기부터 8주간에 걸쳐 실험을 수행 하였고 6개 지점을 실험 point로 설정하였다. 실험방법은 시료의 보관조건 $-20^{\circ}C$로 보관하여 냉동시킨 후 실온에 꺼내어 녹인 다음 $4^{\circ}C$에서 보관 하였다. 그리고 실험항목은 엔도톡신, 미생물한도, HPLC순도, 농도 등의 실험을 진행하였다. 실험결과 엔도톡신은 초기 0.5 EU/mg이하에서 최종 8주차에서도 0.5 EU/mg이하의 결과를 보였으며, 미생물한도에서도 불검출로 나타났고 순도는 210 nm에서 99.23~99.90%로 RSD% 0.23%, 280 nm에서는 100%의 결과를 나타내어 다른 물질로의 변화나 물질 분해특성이 없는 것으로 나타났다. 그리고 농도에서는 최초 55.15 mg/mL에서 최종 55.55 mg/mL로 RSD% 0.55%로 안정된 결과임을 알 수 있었다. 상기 의 실험결과로 rProtein A의 $4^{\circ}C$에서의 8주간의 보관조건은 미생물 발생이나 물질분해 특성이 없이 안정성이 확보 되었다는 결론을 얻을 수 있었다.

• Applied Microscopy
• /
• 제27권4호
• /
• pp.403-416
• /
• 1997

In order to observe the localization of the specific antigenic protein in the tissue of Paragonimus westermani metacercaria, immunogoldlabeling method was applied using IgG of the dog which were infected with Paragonimus westermani metacercaria and IgG of rabbits which were immunized with purified 23 kDa protein from metacercaria of the Paragenimus westermani. The metacercaria worm tissues obtained from Cambaroides similis were embedded in Lowicryl HM20 medium, treated with infected and immunized IgG and protein A gold complex (particle size; 12 nm) and observed by electron microscope. In the tissue antigen of Paragonimus westermani metacercaria, the content of excretory bladder which was highly dense electron density was constituted in the excretory bladder of the parenchymal tissue. In the metacercaria tissues antigen reacted with IgG of infected dog. Labeled gold particles distributed on the interstitial matrix of parenchymal cells, fibrous granules of parenchymal tissue and the content of excretory bladder. High antigenicity was observed on content of excretory bladder. It was found to be specifically distributed at the tissue of Paragonimus westermani metacercaria. In the tissues antigen reacted with IgG of immunized rabbit. Labeled gold particles randomly distributed on the interstitial matrix and fibrous granules of parenchymal tissue but in the content of excretory bladder of Paragonimus westermani metacercaria, gold particles were richly labeled. Therefore, the 23 kDa protein contained with Paragonimus westermani metacercaria was found protein which was specifically constituted at the content of excretory bladder of Paragonimum westermani metacercaria. The 23 kDa protein was commonly contained from of Paragonimus westermani metacercaria to adult and showed strong antigenicity against the immunized and infected IgG.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권4호
• /
• pp.1667-1674
• /
• 2012

Objective: To explore the differential protein expression profile in EC304 gastric cancer cells induced by alphastatin. Methods: Cultured EC304 cells in the exponential phase of growth were randomly divided into alphastatin and control groups. Total proteins were extracted and the two dimensional electrophoresis (2-DE) technique was applied to analyze differences in expression with ImageMaster 2D Platinum 5.0 software. Proteins were identified using the MASCOT database and selected differently expressed proteins were characterised by western blotting and immunofluorescence. Results: $1350{\pm}90$ protein spots were detected by the ImageMaster software in the 2-DE gel images from the control and alphastatin groups. The match rate was about 72-80% for the spectrum profiles, with 29 significantly different protein spots being identified, 10 upregulated, 16 downregulated, two new and one lost. The MASCOT search scores were 64-666 and the peptide matching numbers were 3-27 with sequence coverage of 8-62%. Twenty-three proteins were checked by mass spectrometry, including decrease in Nm23 and profilin-2 isoform b associated with the regulation of actin multimerisation induced by extracellular signals. Conclusion: The proteome in EC304 cells is dramatically altered by alphastatin, which appears to play an important role in modulating cellular activity and anti-angiogenesis by regulating protein expression and signal transduction pathways through Nm23 and profilin-2 isoform b, providing new research directions for anti-angiogenic therapy of gastric cancer.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권8호
• /
• pp.3967-3971
• /
• 2012

Aims: To explore the relationship between various molecular makers and liver metastasis of colorectal cancer (CRC). Method: Using immunohistochemistry, protein expression of CEA, nm23, c-met, MMP2, COX-2, VEGF, EGFR, and CD44 was assessed in 80 CRC cases. The Chi-square test and logistic regression were performed to analyze the relationship between these indicators and CRC liver metastasis. Results: There were significant differences in expression of CEA, MMP2, CD44, VEGF and EGFR between the liver metastasis and non metastasis groups (P < 0.05); no significant differences were noted for nm23, c-met, and COX-2 expression. Logistic regression analysis showed that only CEA, VEGF, and EGFR entered into the regression equation, and had significant correlations with CRC liver metastasis (${\alpha}$ inclusion= 0.10, ${\alpha}$ elimination = 0.15, R2 = 0.718). Conclusions: Combination detection of CEA, VEGF, and EGFR may be an effective means to predict CRC liver metastasis. Nm23, c-met, MMP2, COX-2, and CD44, in contrast, are not suitable as prognostic markers.

• 한국물환경학회지
• /
• 제23권6호
• /
• pp.863-871
• /
• 2007

Spectroscopic characteristics of river water from four major watersheds in the Ulsan area were measured to examine their potential for estimating water quality parameters. The total 176 river samples were collected from 44 sites of small streams within the watersheds during the year 2006. Spectroscopic characteristics investigated included protein-like fluorescence (FLF) intensity, fulvic-like fluorescence (FLF) intensity, terrestrial humic-like fluorescence (TLF) intensity, UV absorbance at 254 nm, and UV absorbance difference at 220 nm and 254 nm. Protein-like fluorescence intensity showed linear relationships with biochemical oxygen demand (BOD), chemical oxygen demand (COD), total phosphorous (TP) concentrations of the samples with the correlation of 0.784, 0.779, and 0.733, respectively. Due to the UV absorption characteristics of nitrate at 220 nm, UV absorbance difference at 220 nm and 254 nm was selected to represent total nitrogen (TN) concentration. Exclusion of some samples with PLF intensity higher than 5.0 improved the correlation between the UV absorbance difference and TN as demonstrated by the increase of the correlation coefficient from 0.392 to 0.784. Instead, for the samples with PLF intensity lower than 5.0, the highest correlation of TN was achieved with UV absorbance at 254 nm. The results suggest that PLF intensity could be used as the estimation index for BOD, COD, and TP concentration of river water, and as the primary screening index for the prediction of TN using UV absorbance difference. Some BOD-based water quality levels among the river water were statistically discriminated by the PLF intensity. Low p-values were obtained from the t-tests on the samples with the first level and the second level (p=0.0003) and the samples with the second and the third levels (p=0.0413). Our combined results demonstrated that the selected spectroscopic characteristics of river water could be utilized as a tool for on-site real-time monitoring and/or the primary estimation of water quality.

• 미생물학회지
• /
• 제23권2호
• /
• pp.129-137
• /
• 1985

The RK-temperate phage which infected with Bacillus cereus was isolated and the characters were investigated. The induction of RK-temperate phage from host bacterium attained by ultraviolet light irradiation (15W, 30cm, 30-120sec) and mitomycin C treatment (0.2-2 ug/ml). The host range of RK-temperate phage was not revealed with lysogenic and related strains of B. cereus. But B. cereus(PS) 352 which obtained by N-nitrosoguanidine treatment (1,000{$\mu}g/ml)$ to phage infected with host bacteria was sensitive bacteria of RK-temperate phage. RK-temperate phage was stabilized at the condition of nutrient broth (pH 7-8), Tris-buffer (pH 7-8) and ammonium buffer (pH 8-9) and Sorensen's phosphate buffer (pH 6-7), but unstabilized at other salt solutions and pH range. Also, thermostability was to $45^{\circ}C$ but unstabilized at above $50^{\circ}C$. At RK-temperate phage, the measurment values of head, neck, mid tail and end tail were 59nm, $9{\times}16nm,\;10{\times}189nm,\;and\;10{\times}14nm$ respectively. The morphology of head was regular polyhedron, and the end tail was coneate form. On the one hand, the number of capsid protein layer of tail were consist of 4, 35, and 1 at neck, mid tail, and end tail, respectively. RK-temperate phage was identified with DNA phage and G+C contents were 38.63. The latent time of RK-temperate phage was 30 minutes and the burst size was 70-80. And the host bacteria was lysed in case of multi-infection, above moi 1.

• Asian-Australasian Journal of Animal Sciences
• /
• 제12권6호
• /
• pp.891-894
• /
• 1999

A field experiment was conducted with Calliandra calothyrsus of fifteen provenances to evaluate the variation in the condensed tannin content of fresh leaves. Plants were raised in a nursery for three months and planted in the field. Experimental design was line planting of double hedgerows (3 m long plot, 1/2 meter interval, 45 plants in each plot) with 5 replicates. Plants were lopped every 5 months and samples (10 different plants within a plot; 4 newly flushed leaves, 4 partially expanded leaves and 4 mature full expanded leaves from each of 10 trees in a plot) were collected and were put on ice before transfer to the laboratory. Proanthocyanidine was extracted in aqueous acetone, and total extractable proanthcyanidine (TEPA) and total proanthocyanidine (TOPA) were measured. In addition, proximate analyses were made of different provenances. Crude protein contents varied from 19.74% to 16.45%; in vitro organic matter digestibility ranged from 36.09% to 23.32%. TEPA content ranged from 11.22 to 16.01 (measured as absorbance at $550nm,\;g^{-1}$ DM) and TOPA from 24.57 to 31.72 (measured as absorbance at $550nm,\;g^{-1}$ DM) TEPA and TOPA had a positive correlation, whereas crude protein content of Calliandra was negatively correlated with TEPA and TOPA. Provenances, such as Georgesville (Belize), local, Coban (Guatemala), Patulul (Guatemala), Bombana (Mexico) can be selected on the basis of nutritive value of fodder.