• Molecules and Cells
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• 제25권3호
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• pp.446-451
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• 2008

We assessed heterologous protein expression in 64 strains obtained from the Escherichia coli Reference (ECOR) collection, a collection representing diverse natural E. coli populations. A plasmid generating a glutathione S-transferase and plant carbonic anhydrase fusion protein (GST-CA) under the control of the tac promoter was introduced into the ECOR strains, and the quantity of the fusion protein was determined by SDS-PAGE. The foreign protein was generated at various levels, from very high (40 strains, high producers) to very low (six strains, low producers). Immunoblotting showed that the high producers expressed approximately 250-500 times more GST-CA protein than the low producers. The results of semi-quantitative RT-PCR showed that the low producers generated mRNA levels comparable to those of the high producers, thereby suggesting that, at least in this case, inefficient translation is a major cause of the low production. We introduced a different plasmid, which expressed a maltose binding protein and plant guanylate kinase fusion protein (MBP-GK) into the six low producers. Interestingly, five of these expressed MBP-GK at very high levels. Thus, we conclude that the production of a particular protein from an expression vector can vary considerably, depending on the host strain. Strains in the ECOR collection could function as useful alternative hosts when a desired level of protein expression is not obtained from commonly used strains, such as E. coli K12 or B derivatives.

• Asian-Australasian Journal of Animal Sciences
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• 제15권8호
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• pp.1210-1214
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• 2002

The large and rapid changes of glucose utilization in lactating mammary tissue in response to changes in nutritional state must be largely related by external signal of insulin. This also must be related with the quantity and composition of the diet in vivo. To characterize the mode of gut extract protein with insulin, in vitro experiment was conducted with HC11 cells. The gut extract protein has not only the same effect as insulin alone but also the synergistic effect with insulin in 2-Deoxy[3H] glucose uptake. Although the gut extract did not modulates glucose uptake via increasing the rate of translation of the GLUT1 protein, northern blot analysis indicated that the gut extract protein increased the expression of GLUT1 mRNA by a threefold and also there was a dose-dependent increase in the expression of GLUT1 mRNA. The gut extract protein is therefore shown to be capable of modulating glucose uptake by transcription level with insulin in HC 11 cells.

• Journal of Microbiology and Biotechnology
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• 제30권6호
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• pp.830-838
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• 2020

The histone-like nucleoid structuring protein (H-NS) is an abundant global regulator of environmentally controlled gene expression. Herein, we demonstrate that H-NS represses the expression of LeuO, the master regulator of the cyclic(Phe-Pro)-dependent signaling pathway, by directly binding to the upstream region of the gene. H-NS binds to a long stretched region (more than 160-bp long), which overlaps with binding sites for ToxR and LeuO. A high quantity of H-NS outcompetes ToxR for binding to the cis-acting element of leuO. However, our footprinting analyses suggests that the binding of H-NS is relatively weaker than LeuO or ToxR at the same molarity. Considering that the DNA nucleotide sequences of the upstream regions of leuO genes are highly conserved among various Vibrio, such patterns as those found in V. vulnificus would be a common feature in the regulation of leuO gene expression in Vibrionaceae. Taken together, these results suggest that, in species belonging to Vibrionaceae, H-NS regulates the expression of leuO as a basal stopper when cFP-ToxR mediated signaling is absent.

• 한국건축시공학회:학술대회논문집
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• 한국건축시공학회 2013년도 춘계 학술논문 발표대회
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• pp.210-212
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• 2013

The construction cost estimates during the design phase becomes the standard to judge profitability and validity, and is very important in various decision-makings by project owner. However, since approximate costs are quoted when many parts are undecided in the early stage of project, differences are bound to occur between the construction cost calculated through approximate quotation and that put into construction actually. Also, since in existing quotation works, quantity calculations have been dependent on the staff's manual work, involving error potential, and thus differences are likely in quantity calculation depending on the quotation staff's method of calculation. In this study, the process of creating space model to deduce 3D geometry information for approximate quotation in association with knowledge information and the expression for calculation of finishing area were proposed.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1620-1627
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• 2009

The design and expression of an antihypertensive peptide multimer (AHPM), a common precursor of 11 kinds of antihypertensive peptides (AHPs) tandemly linked up according to the restriction sites of gastrointestinal proteases, were explored. The DNA fragment encoding the AHPM was chemically synthesized and cloned into expression vector pGEX-3X. After an optimum induction with IPTG, the recombinant AHPM fused with glutathione S-transferase (GST-AHPM) was expressed mostly as inclusion body in Escherichia coli BL21 and reached the maximal production, 35% of total intracellular protein. The inclusion body was washed, dissolved, and purified by cation-exchange chromatography under denaturing conditions, followed by refolding together with size-exclusion chromatography and gradual dialysis. The resulting yield of the soluble GSTAHPM (34 kDa) with a purity of 95% reached 399 mg/l culture. The release of high active fragments from the AHPM was confirmed by the simulated gastrointestinal digestion. The results suggest that the design strategy and production method of the AHPM will be useful to obtain a large quantity of recombinant AHPs at a low cost.

• 인터넷정보학회논문지
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• 제6권1호
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• pp.85-93
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• 2005

본 논문은 다량의 고차원 얼굴 표정 모션 데이터를 2차원 공간에 분포시키고, 애니메이터가 이 공간을 항해하면서 원하는 표정들을 실시간 적으로 선택함으로써 얼굴 표정 애니메이션을 생성하는 방법을 기술한다. 본 논문에서는 약 2400여개의 얼굴 표정 프레임을 이용하여 표정공간을 구성하였다. 표정공간의 생성은 임의의 두 표정간의 최단거리의 결정으로 귀결된다. 표정공간은 다양체 공간으로서 이 공간내의 두 점간의 거리는 다음과 같이 근사적으로 표현한다. 임의의 마커간의 거리를 표시하는 거리행렬을 사용하여 각 표정의 상태를 표현하는 표정상태벡터를 정의한 후, 두 표정이 인접해 있으면, 이를 두 표정 간 최단거리(다양체 거리)에 대한 근사치로 간주한다. 그리하여 인접 표정들 간의 인접거리가 결정되면, 이들 인접거리들을 연결하여 임의의 두 표정 상태간의 최단거리를 구하는데, 이를 위해 Floyd 알고리즘을 이용한다. 다차원 공간인 표정공간을 가시화하기 위해서는 CCA 투영기법을 이용하여 2차원 평면에 투영시켰다 얼굴 애니메이션은 사용자 인터베이스를 사용하여 애니메이터들이 2차원 공간을 항해하면서 실시간으로 생성한다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권6호
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• pp.767-771
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• 2005

In order to understand the molecular basis of chicken heterosis in reproduction traits, mRNA differential display (DDRT-PCR) methods were used to analyze the differential gene expression of ovary tissue between hybrids and their parental lines in a 4${\times}$4 diallel cross, involving 4 chicken breeds, which were White Plymouth Rock (E), CAU Brown (D), Silkies (C) and White Leghorn (A). Total of 331 differential displayed cDNA bands from 1,161 were displayed in the 4${\times}$4 diallel cross combinations with 30 pairs of primers, which shows the differences of gene expression between hybrids and their parental lines were very obvious in quantity and quality. Seven types of differential expression patterns were found: Co-dominance expressed pattern (T1), under-expression of parental fragments in hybrids (T2), over-expression of parental fragments in hybrids (T3), hybrid-absence expressed pattern (T4), single parentspecific expressed pattern (T5), dominant expression fragments of single parent in hybrids (T6), hybrid-specific expressed pattern (T7). Correlation analysis indicated that there were significant correlations between the pattern of T3 and the heterosis percentage of egg number of 32-week and 42-week old chickens(p<0.01), while there were negative significant correlations between the pattern of T7 and the heterosis percentage of egg number of 32-week and 42 week-old birds (p<0.01).

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6595-6599
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• 2013

Leptin and its receptor are involved in breast carcinogenesis as mitogenic factors. Therefore, they could be considered as targets for breast cancer therapy. Expression of the leptin receptor gene could be modulated by leptin secretion. Silibinin and curcumin are herbal compounds with anti-cancer activity against breast cancer. The aim of this study was to assess their potential to inhibit of expression of the leptin gene and its receptor and leptin secretion. Cytotoxic effects of the two agents on combination on T47D breast cancer cells was investigated by MTT assay test after 24h treatment. With different concentrations the levels of leptin, leptin receptor genes expression were measured by reverse-transcription real-time PCR. Amount of secreted leptin in the culture medium was determined by ELISA. Data were statistically analyzed by one-way ANOVA test. The silibinin and curcumin combination inhibited growth of T47D cells in a dose dependent manner. There were also significant difference between control and treated cells in leptin expression and the quantity of secreted leptin with a relative decrease in leptin receptor expression. In conclusion, these herbal compounds inhibit the expression and secretion of leptin and it could probably be used as drug candidates for breast cancer therapy through leptin targeting in the future.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.639-643
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• 2013

Aim: Connexin 43 (Cx43) and E-cadherin are important biomarkers related with cancer. Their expression at protein and mRNA levels was here investigated in 50 primary lung carcinoma tissues and 20 samples of adjacent normal tissue of Chinese patients with non-small cell lung cancer (NSCLC). Methods: Protein and mRNA expression were evaluated by ABC immunohistochemistry and RT-PCR. Results: (1) The positive expression rates of Cx43 and E-cadherin protein were higher in the adjacent normal tissues than those in the primary lung carcinoma tissues; (2) the positive expression rates of Cx43 and E-cadherin protein decreased with NSCLC progression; (3) the expression of E-cadherin protein was not related with the pathological type of NSCLC; and (4) the relative quantity of the Cx43 or E-cadherin mRNA expression was correlated with the the histological type, clinical stage, cancer cell differentiation and the lymph node metastasis. Conclusion: The data suggested that the Cx43 and E-cadherin are reduced with NSCLC progression, and might be important biomarkers for judging the metastasis and prognosis.

• Journal of Animal Science and Technology
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• 제63권6호
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• pp.1397-1410
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• 2021

The present study was designed to determine the influence of all-trans retinoic acid (ATRA) on adipogenesis-related gene regulation in bovine intramuscular (IM) and subcutaneous (SC) adipose cells during differentiation. Bovine IM and SC adipocytes were isolated from three 19-mo-old, crossbred steers. Adipogenic differentiation was induced upon cultured IM and SC preadipocytes with various doses (0, 0.001, 0.01, 0.1, 1 µM) of ATRA. After 96 h of incubation, cells were harvested and used to measure the gene expression of CCAAT/Enhancer binding protein β (C/EBPβ), peroxisome proliferator-activated receptor (PPAR) γ, glucose transporter 4 (GLUT4), stearoyl CoA desaturase (SCD), and Smad transcription factor 3 (Smad3) relative to the quantity of ribosomal protein subunit 9 (RPS 9). Retinoic acid receptor (RAR) antagonist also tested to identify the effect of ATRA on PPARγ -RAR related gene expression in IM cells. The addition of ATRA to bovine IM decreased (p < 0.05) expression of PPARγ. The expression of PPARγ was also tended to be downregulated (p < 0.1) in high levels (10 µM) of ATRA treatment in SC cells. The treatment of RAR antagonist increased the expression of PPARγ in IM cells. Expression of C/EBPβ decreased (p < 0.05) in SC, but no change was observed in IM (p > 0.05). Increasing levels of ATRA may block adipogenic differentiation via transcriptional regulation of PPARγ. The efficacy of ATRA treatment in adipose cells may vary depending on the location.