• Title/Summary/Keyword: total protein expression

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Production of the taste-modifying protein, miraculin, in transgenic lettuce

  • Ezura, Hiroshi;Sun, Heyon-Jin
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.126-131
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    • 2005
  • Richadella dulcifica, a native shrub in tropical West Africa, gives red berries that have the unusual property of modifying a sour taste into a sweet taste. The red berries contain a taste-modifying protein named miraculin. A synthetic gene encoding miraculin was placed under the control of constitutive promoters and transferred to lettuce. High expression of miraculin was obtained, with accumulation of up to 1% total soluble protein in lettuce leaf. In addition, the miraculin expressed in lettuce possesses a taste-modifying activity.

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The Effect of Exercise Training on Blood and Metabolic Variances, and Genes Expressions in Hyperlipidemic Rats (고지혈증 흰쥐에서 운동훈련이 혈액, 대사 변인 및 유전자 발현에 미치는 영향)

  • Kim, Kihoon;Kwon, Taedong;Kim, Jongyeon
    • Korean Journal of Exercise Nutrition
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    • v.13 no.2
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    • pp.93-100
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    • 2009
  • This study was conducted to investigate the effect of exercise training on blood and metabolic variances and genes expressions in hyperlipidemic rats. Three weeks-old male rats were randomly assigned into chow (n=7), high-fat diet (HF, n=7) and HF+exercise (HF+EX, n=7) groups. Exercise training consisted of the treadmill running 5 times per week during 8 weeks (0% grade, 30 min/time for first 4 weeks and 0% grade, 60 min/time the other 4weeks). The levels of triglyceride and total -cholesterol were increased in HF diet compared with chow group, and recovered to level of chow group by exercise training. Plasma glucose and insulin concentrations increased by 40 and 50%, respectively in HF diet compared with chow diet group, and these increases returned to the level of chow group by exercise training (p<.05). Body weight and abdominal fat mass were increased by high-fat diet compared with chow diet, and recovered to level of chow group by exercise training. Long-chain fatty acid oxidation rate and AMPK protein expression was not changed by HF diet, but increased by exercise training compared with high-fat diet (p<.05). UCP3 protein expression was not changed by either high-fat diet or exercise training compared with chow group. There was high correlation between plasma triglyceride and total cholesterol concentrations(p<.01). Plasma triglyceride or total cholesterol level showed correlation with following factors; plasma insulin and glucose levels, body weight, abdominal fat weight, UCP3 protein expression and long-chain fatty acid oxidation rate. These results showed that exercise training on the treadmill recovered hyperlipidemia, hyperglycemia and hyperinsulinemia induced by high-fat diet for 8 weeks. These exercise effects may be related with decreased body weight and abdominal fat mass, and increased long-chain fatty acid oxidation rate.

A SERI technique reveals an immunosuppressive activity of a serine-rich protein encoded in Cotesia plutellae bracovirus

  • Barandoc, Karen P.;Park, Jay-Young;Kim, Yong-Gyun
    • BMB Reports
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    • v.43 no.4
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    • pp.279-283
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    • 2010
  • Polydnavirus genome is segmented and dispersed on host wasp chromosome. After replication, the segments form double- stranded circular DNAs and embedded in viral coat proteins. These viral particles are delivered into a parasitized host along with parasitoid eggs. A serine-rich protein (SRP) is predicted in a polydnavirus, Cotesia plutellae bracovirus (CpBV), genome in its segment no. 33 (CpBV-S33), creating CpBV-SRP1. This study explored its expression and physiological function in the diamondback moth, Plutella xylostella, larvae parasitized by C. plutellae. CpBV-SRP1 encodes 122 amino acids with 26 serines and several predicted phosphorylation sites. It is persistently expressed in all tested tissues of parasitized P. xylostella including hemocyte, fat body, and gut. Its physiological function was analyzed by injecting CpBV-S33 and inducing its expression in nonparasitized P. xylostella by a technique called SERI (segment expression and RNA interference). The expression of CpBV-SRP1 significantly impaired the spreading behavior and total cell count of hemocytes of treated larvae. Subsequent RNA interference of CpBV-SRP1 rescued the immunosuppressive response. This study reports the persistent expression of CpBV-SRP1 in a parasitized host and its parasitic role in suppressing the host immune response by altering hemocyte behavior and survival.

High-Level Constitutive Expression of Mouse CD4 and CD4/CD8${\alpha}$ Hybrid Molecules in Transgenic Mice

  • Kim, Joongkyu;Choi, Young-Il;Park, Sang-D;Seong, Rho-H
    • Animal cells and systems
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    • v.1 no.4
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    • pp.657-663
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    • 1997
  • The CD4 and CDS coreceptors, in conjunction with the T cell receptor (TCR) , make important contributions to the differentiation of thymocytes. They have been shown to be involved in the clonal deletion and positive selection processes during T cell development in thymus. To further analyze the role of CD4 and CDS proteins during T cell differentiation, we have generated transgenic mice constitutively expressing high levels of a native CD4 and a CD4{CDSa hybrid protein. The hybrid protein is composed of CD4 extracellular domain linked to the CD8a transmembrane region and cytoplasmic tail. The transgenes were driven by human beta-actin promoter, and therefore, they were expressed in all tissues examined including thymus, spleen, and lymph nodes. The resulting CD4 and CD4{CD8${\alpha}$transgenic mice were found to express the CD4 and CD4{CD8${\alpha}$ respectively, in developing thymocytes and peripheral T cells. The expression levels of transgenic proteins were 5-10 times higher than that of endogenous CD4 in thymus. However, total surface CD4 expression (CD4 or CD4{CD8${\alpha}$ transgenic protein plus endogenous CD4) of the transgenic mice were main. tained at similar levels compared to control littermates. Surface CD4 expression on CDS T cells, however, was significantly lower than that on cells expressing endogenous CD4. These results suggest that a total avidity between developing thymocytes and thymic stromal cells is impor. tant for differentiation of thymocytes.

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The Role of Protein Kinase C and Protein Tyrosine Kinase in the Signal Transduction Pathway of Stimulus Induced by Endotoxin in Peripheral Blood Monocyte (말초혈액 단핵구에 대한 내독소 자극의 신호 전달에서 Protein Kinase C와 Protein Tyrosine Kinase의 역할)

  • Kim, Jae-Yeol;Park, Jae-Suk;Lee, Gwi-Lae;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.2
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    • pp.338-348
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    • 1997
  • Background : Endotoxin, the component of outermembrane of gram negative organism, plays an important role in the initiation and amplification of inflammatory reaction by its effects on inflammatory cells. Until recently, there have been continuing efforts to delinate the mechanisms of the signal trasduction pathway of endotoxin stimuli on inflammatory cells. By uncovering the mechanisms of signal transduction pathway of endotoxin stimuli, we can expect to have tools to control the excessive inflammatory responses which sometimes may be fatal to the involved host. It was generally accepted that endotoxin exerts its inflammatory effects through inflammatory cytokines that are produced by endotoxin-stimulated inflammatory cells and there were some reports on the importance of protein kinase C and protein tyrosine kinase activation in the production of inflammatory cytokines by endotoxin So we evaluated the effect of pretreatment of protein kinase C inhibitors (H7, Staurosporin) and protein tyrosine kinase inhibitors(Herbimycin, Genistein) on the endotoxin-stimulated cytokines(IL-8 & TNF-$\alpha$) mRNA expression. Method : Peripheral blood monocytes were isolated from healthy volunteers by Ficoll-Hypaque density gradient method and purified by adhesion to 60mm Petri dishes. Endotoxin(LPS 100ng/ml) was added to each dishes except one control dish, and each endotoxin-stimulated dishes was preincubated with H7, Staurosporin(protein kinase C inhibitor), Herbimycin or Genistein(protein tyrosine kinase inhibitor) respectively except one dish. Four hours later the endotoxin stimulation, total RNA was extracted and Northern blot analysis for IL-8 mRNA and TNF-$\alpha$ mRNA was done. Result : Endotoxin stimulation increased the expression of IL-8 mRNA and TNF-$\alpha$ mRNA expression in human peripheral blood monocyte as expected and the stimulatory effect of endotoxin on TNF-$\alpha$ mRNA expression was inhibited by protein kinase C inhibitors(H7, Staurosporin) and protein tyrosine kinase inhibitors (Herbimycin, Genistein). The inhibitory effect of each drugs was increased with increasing concentration. The stimulatory effect of endotoxin on IL-8 mRNA was also inhibited by H7 and protein tyrosine kinase inhibitors (Herbimycin, Genistein) dose-dependently but not by Staurosporin. Conclusion : Protein kinase C and protein tyrosine kinase are involved in the endotoxin induced signal transduction pathway in human peripheral blood monocyte.

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An Iterative Spot Matching for 2-Dimensional Protein Separation Images (반복 점진적 방법에 의한 2차원 단백질 분리 영상의 반점 정합)

  • Kim, Jung-Ja;Hoang, Minh T.;Kim, Dong-Wook;Kim, Nam-Gyun;Won, Yong-Gwan
    • Journal of Biomedical Engineering Research
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    • v.28 no.5
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    • pp.601-608
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    • 2007
  • 2 Dimensional Gel Electrophoresis(2DGE) is an essentialmethodology for analysis on the expression of various proteins. For example, information for the location, mass, expression, size and shape of the proteins obtained by 2DGE can be used for diagnosis, prognosis and biological progress by comparison of patients with the normal persons. Protein spot matching for this purpose is comparative analysis of protein expression pattern for the 2DGE images generated under different conditions. However, visual analysis of protein spots which are more than several hundreds included in a 2DGE image requires long time and heavy effort. Furthermore, geometrical distortion makes the spot matching for the same protein harder. In this paper, an iterative algorithm is introduced for more efficient spot matching. Proposed method is first performing global matching step, which reduces the geometrical difference between the landmarks and the spot to be matched. Thus, movement for a spot is defined by a weighted sum of the movement of the landmark spots. Weight for the summation is defined by the inverse of the distance from the spots to the landmarks. This movement is iteratively performed until the total sum of the difference between the corresponding landmarks is larger than a pre-selected value. Due to local distortion generally occurred in 2DGE images, there are many regions in whichmany spot pairs are miss-matched. In the second stage, the same spot matching algorithm is applied to such local regions with the additional landmarks for those regions. In other words, the same method is applied with the expanded landmark set to which additional landmarks are added. Our proposed algorithm for spot matching empirically proved reliable analysis of protein separation image by producing higher accuracy.

Gene Expression Profiling of Non-Hodgkin Lymphomas

  • Zekri, Abdel-Rahman Nabawy;Hassan, Zeinab Korany;Bahnassy, Abeer Ahmed;Eldahshan, Dina Hassan;El-Rouby, Mahmoud Nour Eldin;Kamel, Mahmoud Mohamed;Hafez, Mohamed Mahmoud
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4393-4398
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    • 2013
  • Background: Chromosomal translocations are genetic aberrations associated with specific non-Hodgkin lymphoma (NHL) subtypes. This study investigated the differential gene expression profile of Egyptian NHL cases based on a microarray approach. Materials and Methods: The study included tissue samples from 40 NHL patients and 20 normal lymph nodes used as controls. Total RNA was extracted and used for cDNA microarray assays. The quantitative real time polymerase chain reaction was used to identify the aberrantly expressed genes in cancer. Results: Significant associations of 8 up-regulated and 4 down-regulated genes with NHL were observed. Aberrant expression of a new group of genes not reported previously was apparent, including down-regulated NAG14 protein, 3 beta hydroxy-delta 5-c27 steroid oxi-reductase, oxi-glutarate dehydrogenase (lipo-amide), immunoglobulin lambda like polypeptide 3, protein kinase x linked, Hmt1, and caveolin 2 Tetra protein. The up-regulated genes were Rb binding protein 5, DKFZP586J1624 protein, protein kinase inhibitor gamma, zinc finger protein 3, choline ethanolamine phospho-transferase CEPT1, protein phosphatase, and histone deacetylase-3. Conclusions: This study revealed that new differentially expressed genes that may be markers for NHL patients and individuals who are at high risk for cancer development.

Design and Expression of High Nutritional Peptide (HEAAE) in E. coli

  • Kim, Jae-Ho;Lee, Chang-Kook;Hong, Bum-Shik
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.132-137
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    • 1997
  • A novel protein (HEAAE, High Essential Amino Acid Encoding Protein), rich in essential amino acids ($75{\%}$ of total), was designed and constructed in our laboratory. The designed peptides were analyzed by SYBLE and stable secondary and tertiary structures were predicted. The monomeric form (HEAAE-1) of the protein consists of 20 amino acid residues with four additional amino acids comprising a potential ${\beta}$-turn (HEAAE-4). Size exclusion analysis demonstrated that the monomer is self-aggregates in aqueous solution to form higher ordered multimeric structures, which are very reminiscent of natural plant storage proteins. The DNA encoding this amino acid sequence was synthesized, and from this monomeric gene fragment (heaae-1), the stable tetrameric form of the gene (heaae-4) was generated by subcloning into the E. coli expression vector pKK223-3. A clear 6 kDa polypeptide band corresponding to the molecular weight of the dimeric form (HEAAE-2) was detected. The smeared band which appeared around the molecular weight corresponding to HEAAE-4 of 11 kDa suggested that the tetramer form of this protein might be processed into smaller size products.

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Peanut sprouts extract (Arachis hypogaea L.) has anti-obesity effects by controlling the protein expressions of $PPAR{\gamma}$ and adiponectin of adipose tissue in rats fed high-fat diet

  • Kang, Nam E;Ha, Ae Wha;Woo, Hye Won;Kim, Woo Kyoung
    • Nutrition Research and Practice
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    • v.8 no.2
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    • pp.158-164
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    • 2014
  • BACKGROUD/OBEJECTIVES: This study aims to find out the effects of peanut sprout extracts on weight controls and protein expressions of transcription factors related to adipocyte differentiation and adipocytokine in rats under high-fat diets. MATERIALS/METHODS: Four week-old Sparague-Dawley (SD) were assigned to 4 groups; normal-fat (NF) diets (7% fat diet), high-fat (HF) diets (20% fat diet), high fat diets with low peanut sprout extract (HF + PSEL) diet (20% fat and 0.025% peanut sprout extract), and high fat diets with high peanut sprout extract (HF + PSEH) diet (20% fat and 0.05% peanut sprout extract). Body weight changes, lipid profiles in adipose tissue, and the mRNA protein expressions, such as peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), CCAAT element binding protein ${\alpha}$ (C/EBP ${\alpha}$), leptin, and adiponectin, were determined. RESULTS: After 9 weeks of feeding, the HF + PSEH group had significantly less weight gains than the HF group (P < 0.05). However, the total dietary intakes or food efficiency ratios among groups were not significantly different. The weight of epididymal fat in HF + PSEH group, $3.61{\pm}0.5g$, or HF + PSEL group, $3.80{\pm}0.7g$, was significantly lower than the HF group, $4.39{\pm}0.4g$, (P < 0.05). Total lipids and total cholesterol in adipose tissue were significantly decreased in HF + PSEH group compared to those in the HF group, respectively (P < 0.05). PSEH supplementation caused AST and ALT levels to decrease when it compared to HF group, but it was not statistically significant. The protein expression of $PPAR{\gamma}$ in HF + PSEH group was significantly lower than the HF group (P < 0.05). Comparing with the HF group, the protein expression of adiponectin in HF + PSEH group was significantly increased (P < 0.05). The protein expressions of C/EBP ${\alpha}$ and leptin in HF + PSEH group were lower than the HF group, but it was not statistical significant. CONCLUSIONS: In conclusion, peanut sprout extract has anti-obesity effect by lowering the expressions of $PPAR{\gamma}$ which regulates the expression of adiponectin.

Identification and Expression Patterns of fvexpl1, an Expansin-Like Protein-Encoding Gene, Suggest an Auxiliary Role in the Stipe Morphogenesis of Flammulina velutipes

  • Huang, Qianhui;Han, Xing;Mukhtar, Irum;Gao, Lingling;Huang, Rongmei;Fu, Liping;Yan, Junjie;Tao, Yongxin;Chen, Bingzhi;Xie, Baogui
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.622-629
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    • 2018
  • Expansins are cell wall proteins that mediate cell wall loosening and promote specific tissue and organ morphogenesis in plants and in some microorganisms. Unlike plant expansins, the biological functions of fungal expansin-like proteins have rarely been discussed. In the present study, an expansin-like protein-encoding fvexpl1 gene, was identified from Flammulina velutipes by using local BLAST. It consisted of five exons with a total length of 822 bp. The deduced protein FVEXPL1 contained 274 amino acids with a predicted molecular mass and isoelectric point of 28,589 Da and pH 4.93, respectively. The first 19 amino acids from the N terminal are the signal peptide. Phylogenetic analysis and multiple protein alignment indicated FVEXPL1 was an expansin-like protein. The expression level of fvexpl1 gene in the stipe was significantly higher than that in the mycelia, primordia, and cap. However, the expression level of fvexpl1 gene was significantly higher in the fast elongation region of the stipe as compared with the slow elongation region. Expression analysis indicated that fvexpl1 gene might have an auxiliary role in the stipe morphogenesis of F. velutipes.