• Title/Summary/Keyword: Activity Region

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Identification of Endothelial Specific Region in the Intracellular Adhesion Molecule-2 (ICAM2) Promoter of Miniature Pig

  • Jang, Hoon;Jang, Won-Gu;Kim, Dong Un;Kim, Eun-Jung;Hwang, Sung Soo;Oh, Keon Bong;Lee, Jeong-Woong
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.207-212
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    • 2012
  • The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.

Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2)

  • Lee, Sang-Mi;Kim, Hye-Min;Moon, Seung-Ju;Kang, Man-Jong
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.3
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    • pp.421-427
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    • 2012
  • The production of therapeutic proteins from transgenic animals is one of the most important successes of animal biotechnology. Milk is presently the most mature system for production of therapeutic proteins from a transgenic animal. Specifically, ${\beta}$-casein is a major component of cow, goat and sheep milk, and its promoter has been used to regulate the expression of transgenic genes in the mammary gland of transgenic animals. Here, we cloned the porcine ${\beta}$-casein gene and analyzed the transcriptional activity of the promoter and intron 1 region of the porcine ${\beta}$-casein gene. Sequence inspection of the 5'-flanking region revealed potential DNA elements including SRY, CdxA, AML-a, GATA-3, GATA-1 and C/EBP ${\beta}$. In addition, the first intron of the porcine ${\beta}$-casein gene contained the transcriptional enhancers Oct-1, SRY, YY1, C/EBP ${\beta}$, and AP-1, as well as the retroviral TATA box. We estimated the transcriptional activity for the 5'-proximal region with or without intron 1 of the porcine ${\beta}$-casein gene in HC11 cells stimulated with lactogenic hormones. High transcriptional activity was obtained for the 5'-proximal region with intron 1 of the porcine ${\beta}$-casein gene. The ${\beta}$-casein gene containing the mutant TATA box (CATAAAA) was also cloned from another individual pig. Promoter activity of the luciferase vector containing the mutant TATA box was weaker than the same vector containing the normal TATA box. Taken together, these findings suggest that the transcription of porcine ${\beta}$-casein gene is regulated by lactogenic hormone via intron 1 and promoter containing a mutant TATA box (CATAAAA) has poor porcine ${\beta}$-casein gene activity.

Deletion of N-terminal End Region of ErmSF Leads to an Amino Acid Having Important Role in Methyl Transfer Reaction (ErmSF에서 특이적으로 발견되는 N-terminal End Region의 점차적인 제거에 의한 활성에 중요한 아미노산의 규명)

  • Lee Hak Jin;Jin Hyung Jong
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.257-262
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    • 2004
  • ErmSF is one of the ERM proteins which transfer the methyl group to A2058 in 23S rRNA to confer the resis­tance to MLS (macrolide-lincosamide-streptogramin B) antibiotics on microorganism. Unlike other ERM pro­teins, ErmSF contains long N-terminal end region (NTER), of which $25\%$ is composed of arginine that is known to interact with RNA well. Gradual deletion of NTER leaded us to the point where mutant protein lost much of activity in vivo. Overexpressed and purified mutant protein showed much reduced activity in vitro: $2\%$ activity relative to that of wild type protein. This fact suggests that this amino acid interact with RNA close to meth­ylatable adenine to locate it at an active site properly.

Studies on the Comparative Analysis of Mating Locus (Y-region) of Schizophyllum commune (치마버섯 Mating Locus(Y-region)의 비교분석에 관한 연구)

  • 이인선;박동철
    • Journal of Life Science
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    • v.12 no.2
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    • pp.173-181
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    • 2002
  • This study was conducted to do the comparative analysis of mating type locus controlling the direct formation of fruiting body in Schizophyllum commune which is indigenous to North America with that of other identified mating locus. The 3120 bp Y-region nucleotide of A $\alpha$ 3 mating locus activating a developmental pathway in S. commune was determined, and appeared to have about 96% homology to S. commune 1-71 $A\alpha$3 allele indigenous to South America, showing strongly a conservative feature. This nucleotide analysis also showed above 96% homology highly in the seven presumed exons, and about 97% in the acidic rich region (AR), about 99% in homeodomain (H7), about 97% in the basic rich region (BR), about 95% in the serine rich region (Ser) respectively. In the comparative analysis to the translated polypeptide sequence, S. commune A $\alpha$ 3 mating locus containing Y-region also showed about 97% homology to the region of S. commune indigenous to North America, but the identity ratio to Y1 including Y4, Y5, Y6 different allele types was declined to about 41~49%. In the analysis of functional loci controlling mating activity, it is assumed to have a highly conservative feature showing about 98% homology in homeodomain polypeptide. Especially, it is notable that the homology ratio of above 85% in homeodomain motif between mating type alleles was higher than in the AR, BR, Ser showing about 10~50% homology.

Studied on Garment Restraint(III) - Relation between Clothing Pressure and Muscular Activity of Foundation - (의복의 구속성에 관한 연구(III) - 화운데이션의 의복압과 근활동과의 관계를 중심으로 -)

  • Shim, Boo Ja;Choi, Seon Hee
    • Journal of the Korean Society of Clothing and Textiles
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    • v.17 no.2
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    • pp.197-206
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    • 1993
  • We studied relation between the clothing pressure applied by foundations(waist nipper, girdle, body suit) on the waist of bodies and muscular activity, feeling of tightness. The main results were summerized as follows ; 1. Clothing pressure applied by foundations was high in order of girdle>waist nipper>body suit, also clothing pressure was higher back than front and side, sitting on the chair than standing posture, ventral flection than repose. Individual differences, even if size of body was equal, were shown in clothing pressure applied by foundations with subcutaneous fat's amount in measuring region. 2. The muscular activity of rectus abdominis than obliquus externus abdominis was more affected by foundations in all kinds of postures and motions. Amplitude of electromyogram was high sitting on the chair than standing posture, but there was little difference with motion variation. The wearing girdle strongly affected on the muscular activities of rectus abdominis and obliquus externus abdominis as compared with waist nipper and body suit. 3. The value for feeling of tightness by wearing waist nipper was higher than girdle and body suit. Also the case when sitting on the chair and ventral flection, the value for feeling of tightness was high. When the foundations were on the body, most tightened on the region of the body was anterior abdominal region.

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Gravity wave activities in the polar region using FORMOSAT-3 GPS RO observations

  • Liou, Yuei-An;Yan, Shiang-Kun
    • Proceedings of the KSRS Conference
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    • 2007.10a
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    • pp.65-68
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    • 2007
  • FORMOSAT-3 was launched in April of 2006. It consists of six low earth orbit (LEO) satellites that will be eventually deployed to an orbit at 800 km height. Its scientific goal is to utilize the radio occultation (RO) signals to measure the bending angles when the GPS signals transect the atmosphere. The bending angle is then used to infer atmospheric parameters, including refractivity, temperature, pressure, and relative humidity fields of global distributions through inversion schemes and auxiliary information. The expected number of RO events is around 2500 per day, of which 200 events or so fall into the polar region. Consequently, the FORMOSAT-3 observations are expected to play a key role to improve our knowledge in the weather forecasting and space physics research in the polar region. In this paper, we use temperature profiles retrieved from FORMOSAT-3 RO observations to study the climatology of gravity wave activity in the polar region. FORMOSAT-3 can provide about 200 RO observations a day in the polar region, much more than previous GPS RO missions, and, hence, more detailed climatology of gravity wave activity can be obtained.

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Effects of the trunk stabilization exercise on muscle activity in lumbar region and balance in the patients with hemiplegia (중추신경발달치료를 이용한 몸통 안정화 운동이 뇌졸중 환자의 허리부위 근 활성도와 균형에 미치는 효과)

  • Shim, Hyun-Bo;Cho, Hwi-young;Choi, Won-Ho
    • The Journal of Korean Physical Therapy
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    • v.26 no.1
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    • pp.33-40
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    • 2014
  • Purpose: The aim of this study was to identify the effects of the lower trunk stabilization exercise using neurodevelopmental technique (NDT) on muscle activity in lumbar region and balance in the patients with hemiplegia. Methods: Fifteen participants were allocated in two groups: NDT group (n=8) or control group (n=7). NDT group performed NDT exercise program, while control group conducted walking exercise. Both interventions were given for 30 minutes a day, 3 times a week, for 5 weeks. To measure the muscle activity in rectus abdominis (RA), external oblique (EO) and internal oblique (IO), electromyography (EMG) was used. And, Timed-Up and Go (TUG) test and Berg-Balance Scale (BBS) were performed to assess balance before and after intervention. Results: NDT group showed a significant improvement of muscle activity in RA and EO, while control group did not show significant changes in three muscles. Also, there was a significant difference in muscle activity of RA and EO between two groups. In BBS and TUG test, participants in two groups showed significant improvements after intervention. Especially, significant difference was observed in TUG test between two groups (p<.05). Conclusion: This study demonstrated that NDT exercise is an effective intervention to improve the muscle activity in trunk region and to increase balance in patients with stroke. Thus, we suggested that NDT exercise program would be a treatment intervention in stroke rehabilitation.

Proteolytic Activity of the Crude Enzyme Extracted from the Digestive Tract of Marine Gastropods (해산복족류의 소화관조직중에 분포하는 단백질분해효소의 활성)

  • CHO Deuk Moon;PYEUN Jae Hyeung;BYUN Dae Seok;KIM Chang Yang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.16 no.3
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    • pp.216-224
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    • 1983
  • This work was undertaken to obtain some characteristics of proteolytic enzyme of marine gastropods such as sea hare, Aplysia kurodai, top shell, Turbo cornutus, and abalone, Haliotis discus hannai. An influence of pH, temperature and some chemicals on proteolytic activity of the crude enzyme extracted from digestive tract of the samples was taken into account and the stability of the enzyme during the storage at low temperature was also discussed. In comparison of the activities of the crude enzyme from the samples to the optimum conditions, it was characterized that abalone has twice or the more times higher activities than the other two species of the gastropoda in the acid and weak acid region, while, in the alkaline region, sea hare has six or the more times higher acitivities than the other two species. The proteolytic activity was facilitated by $Mn^{2+}$, some reducing agents, EDTA and DTT, and inhibited by $Hg^{2+}$ and SDS, but the other chemicals were not significantly affected to the activity. The low temperature storage of the enzymes of sea hare and top shell at $0^{\circ}C\;or\;-20^{\circ}C$ was not affected to the enzymic activity under the optimum pH condition except in the alkaline region. On the other hand, the low temperature storage was brought about no significant effect on the activity of the enzymes extracted from abalone under the optimum condition of the weak acid region, but apparently influenced to the activity under the optimum condition of the acid and alkaline region.

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Identification of the+1 Ribosomal Frameshifting Site of LRV1-4 by Mutational Analysis

  • Kim Se Na;Choi Jung Ho;Park Min Woo;Jeong Sun Joo;Han Kyung Sook;Kim Hong Jin
    • Archives of Pharmacal Research
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    • v.28 no.8
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    • pp.956-962
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    • 2005
  • Leishmania virus (LRV)1-4 has been reported to produce a fusion of ORF2 and ORF3 via a programmed +1 frameshift in the region where ORF2 and ORF3 overlap (Lee et a/., 1996). However, the exact frameshift site has not been identified. In this study, we compared the frameshift efficiency of a 259bp (nt. 2565-2823), frameshift region of LRV1-4, and the 71 bp (nt. 2605-2678) sub-region where ORF2 and ORF3 overlap. We then predicted the frameshift site using a new computer program (Pseudoviewer), and finally identified the specific region associated with the mechanism of the LRV1-4's+1 frameshift by means of a mutational analysis based on the predicted structure of LRV1-4 RNA. The predicted structure was confirmed by biochemical analysis. In order to measure the frameshift efficiency, constructs that generate luciferase without a frameshift or with a+1 frameshift, were generated and in vitro transcription/translation analysis was performed. Measurements of the luciferase activity generated, showed that the frameshift efficiency was about $1\%$ for both the 259bp (LRV1-4 259FS) and 71 bp region (LRV1-4 71FS). Luciferase activity was strongly reduced in a mutant (LRV1-4 NH: nt. 2635-2670) with the entire hairpin deleted and in a mutant (LRV1-4 NUS: nt. 2644-2659) with the upper stem of the hairpin deleted. These results indicate that the frameshift site in LRV1-4's is in the 71 bp region where ORF2 and ORF3 overlap, and that nt. 2644-2659 (the upward hairpin stem) playa key role in generating the +1 frameshift.

Secretion of Bacillus subtilis Endo-1,4-$\beta$-D-Glucanase in Yeast Using Promoter and Signal Sequence of Glucoamylase Gene (Glucoamylase 유전자의 promoter 와 분비신호서열을 이용한 Bacillus subtilis Endo-1-4$\beta$-D-Glucanase 의 효모에서 분비)

  • 안종석;강대욱;황인규;박승환;박무영;민태익
    • Korean Journal of Microbiology
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    • v.30 no.5
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    • pp.403-409
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    • 1992
  • For the development of a glucanolytic yeast strain. the seceretion of endo-1.4-$\beta$-D-glucanase (CMCase) of Bacillus subtilis was performed in yeast using glucoamylase gene (STA1) of Saccharomyces diastaticus. A 1.7 kb-DNA fragment of STA1 gene containing authentic promoter, signal sequence, threonine serine-rich (TS) region and N-terminal region (98 amino acids) of mature glucoamylase was ligated to YEp 24. E. coli-yeast shuttle vector. And then. CMCase structural gene of B. subtilis was fused in frame with the 1.7 kb-DNA fragment of STA1 gene, resulting in recombinant plasmid pYES('24. Yeast transformant harboring pYESC24 had no CMCase activity. So. we deleted TS region and N-terminal region of mature glucoamylase existing between signal sequence and CMCase structural gene in pYESC24. consequently constructed recombinant plasmid pYESC11. The yeast transformed with the newly constructed recombinant plasmid pYESC11 efficiently secreted CMCase to extracellular medium. After 4 days culture. total CMCase activity of this transformant was 44.7 units/ml and over 93% of total CMCase activity was detected in culture supernatant.

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