• Title/Summary/Keyword: Functional characterization

Search Result 798, Processing Time 0.031 seconds

Identification of a Novel Gene by EST Clustering and its Expression in Mouse Ovary and Testis (EST Clustering 방법으로 동정한 새로운 유전자의 생쥐 난소 및 정소에서의 발현)

  • Hwang, Sang-Joon;Park, Chang-Eun;Hwang, Kyu-Chan;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.33 no.4
    • /
    • pp.253-263
    • /
    • 2006
  • Objective: Identification of the regulatory mechanism for arrest and initiation of primordial follicular growth is crucial for female fertility. Previously, we found 15 expressed sequence tags (ESTs) that were specifically abundant in the day-S-subtracted cDNA library and that the B357 clone was novel. The present study was conducted to obtain the whole sequence of the novel gene including B357 and to characterize its mRNA and protein expression in mouse ovary and testis. Methods: The extended sequence of the 2,965-bp cDNA fragment for the clone B357 was named ${\underline{5}}-{\underline{d}}ay-{\underline{o}}vary-{\underline{s}}pecific\;gene-{\underline{1}}$ (5DOS1) and submitted to GenBank (accession number ${\underline{AY751521}}$). Expression of 5DOS1 was characterized in both female and male gonads at various developmental stages by Northern blotting, real-time RT-PCR, in situ hybridization, Western blotting, and immunohistochemistry. Results: The 5DOS1 transcript was highly expressed in the adult testis, brain, and muscle as compared to the other tissues. In the ovary, the 5DOS1 transcript was detected in all oocytes from primordial to antral follicles, and highly expressed at day 5 after birth and decreased thereafter. In contrast, expression of 5DOS1 showed a gradual increase during testicular development and its expression was limited to various stages of male germ cells except spermatogonia. Conclusions: This is the first report on the expression and characterization of the 5DOS1 gene in the mouse gonads. Further functional analysis of the 5DOS1 protein will be required to predict its role in gametogenesis.

Functional Characterization of Phosphorylation of the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Nucleocapsid Protein (PRRS 바이러스 Nucleocapsid 단백질 인산화의 기능학적 연구)

  • Lee, Chang-Hee
    • Microbiology and Biotechnology Letters
    • /
    • v.37 no.3
    • /
    • pp.287-292
    • /
    • 2009
  • The nucleocapsid (N) protein of porcine reproductive and respiratory syndrome virus (PRRSV) is a basic multifunctional protein which has been reported to be a serine phosphoprotein with yet-identified functions. As a first step towards understanding the general role of N protein phosphorylation during virus replication, the non-phosphorylated mutant N gene was constructed by mutating all serine residues to alanine. This recombinant N protein was identified to be unphosphorylated, confirming that serine residues truly function as core amino acids responsible for N protein phosphorylation. The PRRSV N protein has been shown to possess the biological features of nuclear localization and N-N homodimerization which individually play critical roles in virus infection. In the present study, therefore, it was attempted to investigate whether these two properties of the N protein are modulated by its phosphorylation status. However, experimental results showed that the non-phosphorylated N protein was still present in the nucleus and nucleolus, and was able to associate with itself by non-covalent interactions. Taken together, the data suggest phosphorylation-independent regulation of N protein nuclear transport or oligomerization, thereby implying the potential involvement of phosphorylation in regulating the activities of the N protein at other levels including RNA-binding capacity.

Improvement on Yield of Extracts from Byproducts of Alaska Pollock Theragra chalcogramma and Sea Tangle Laminaria japonica using Commercial Enzymes and Its Food Component Characterization (상업적 효소를 이용한 명태(Theragra chalcogramma) 두부 및 정형 다시마(Laminaria japonica) 부산물 유래 고압 추출물의 수율개선 및 이의 식품성분 특성)

  • Noh, Yuni;Park, Kwon Hyun;Lee, Ji Sun;Kim, Hyeon Jeong;Kim, Min Ji;Kim, Ki Hyun;Kim, Jeong Gyun;Heu, Min Soo;Kim, Jin-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.46 no.1
    • /
    • pp.37-45
    • /
    • 2013
  • This study was conducted to improve the yield of extracts from Alaska pollock Theragra chalcogramma head and sea tangle Laminaria japonica byproducts using various commercial enzymes, such as Alcalase, Flavourzyme, Neutrase (NH), and Protamex. Among the enzymatic hydrolysates, the yield was highest in hydrolysate incubated with NH for 4 h. NH-treated hydrolysates (NHH) also improved functional properties, such as angiotensin-I converting enzyme (ACE) inhibitory activity and 2,2-diphenyl-1-picryldrazyl (DPPH) radical scavenging activity, as compared to extracts from Alaska pollock head and sea tangle byproducts. Total free amino acid and taste values of NHH were 379.7 mg/100 mL and 24.03, respectively, after digestion for 4 h. These values are 2.2-fold and 1.9-fold higher compared with those of water soluble fractions extracted from Alaska pollock head and non-forming sea tangle, respectively. According to the taste value results, the major taste-active compounds among free amino acids of NHH were glutamic acid and aspartic acid. These results suggest that NHH can be used as an ingredient for natural seasoning preparation.

Chayacterization of Bacillus polyfermenticus SCD as a Probiotic. (Bacillus polyfermenticus SCD의 생균제로서의 특성)

  • 전경동;김혜진;이광호;백현동;강재선
    • Microbiology and Biotechnology Letters
    • /
    • v.30 no.4
    • /
    • pp.359-366
    • /
    • 2002
  • Bacillus polyfermenticus SCD which is commonly called as Bisroot strain is being used for functional foods through the treatment of long-term intestinal disorders, since the live strains in the form of active endospores can successfully reach the target intestine in humans. The cells of B. polyfermenticus SCD were treated for 4h in artificial gastric juice (pH 2.0,3.0) and bile acid. Final viability of the strain in artificial gastric Juice (pH 2.0, 3.0) is reached to 62.8% and 81.2% respectively B. polyfermenticus SCD is resistant to antibiotics such as streptomycin, rifampicin, nystatin and ampicilin. B. polyfermenticus SCD is well known supplies the nutrients by synthesizing vitamin $B_1$, $B_2$, C and K. B. polyfermenticus SCD produces various digestive enzymes and the enzymes enable to completely digest diets in our body. Above all, $\alpha$-amylase and pretense activities are very higher than B. subtilis KCTC 1020, about two fold and twenty five fold respectively. B. polyfermenticus SCD is very stable during long-term storage period in phosphate buffers of wide-range pH, solutions of various concentrations of sodium chloride, 5% glucose solution and water.

Structural and Chemical Characterization of Aquatic Humic Substances in Advanced Water Treatment Processes (고도정수처리 공정에서 수질계 휴믹물질의 구조 및 화학적 특성분석)

  • Kim, Hyun-Chul;Yu, Myong-Jin
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.27 no.3
    • /
    • pp.240-246
    • /
    • 2005
  • Humic substances HS) from process waters at advanced water treatment plant consisted of GAC and Ozone/GAC processes were isolated and extracted by physicochemical fractionation methods to investigate their characteristics. They are characterized for their functionality, chemical composition, spectroscopic characteristics using FT-IR and $^1H$-NMR spectroscopy. Humic fraction gradually decreased from 36.3% to 24.2% from 0.45 to 0.30 mgC/L) through ozonation and carbon adsorption. The humic fraction was isolated into the phenolic and carboxylic groups using A-21 resin, and the concentration of phenolic groups gradually decreased from 38.4% to 23.5% (from 4.9 to $3.2\;{\mu}M/L$ as phenolic-OH) through ozonation and carbon adsorption. In the case of carboxylic groups, the concentration decreased from 61.6% to 43.3% (from 7.8 to $5.8\;{\mu}M/L$ as COOH) through the water treatment processes. On the other hand, concentrations of those roups decreased from 38.4% to 24.0% and 61.6% to 44.9% through carbon adsorption without ozonation, respectively. The structural changes of HS identified from FT-IR and $^1H$-NMR were consistent with the results from the isolation of functional groups in HS.

Synthesis and characterization of hydrophobic and hydrophilic cellulose derivative by esterification (친수성과 소수성을 동시에 가지는 아세틸화 셀룰로스 에테르의 합성 및 특성 평가)

  • Kim, Taehong;Lee, Sangku;Son, Byunghee;Paik, Hyun-Jjong;Yoon, Sanghyeon;Lee, Heesoo
    • Journal of the Korean Crystal Growth and Crystal Technology
    • /
    • v.23 no.1
    • /
    • pp.31-36
    • /
    • 2013
  • Acetylated Cellulose Ether (ACE), cellulose-based amphiphilic polymer with hydrophilic and hydrophobic, was synthesized and investigated in terms of its solubility and wettability for organic solvents and water. Acetyl group was substituted to the cellulose ether in a hydrophilic polymer by esterification. As a result of FT-IR, the peak corresponding to the hydroxyl group decreased and carboxyl acid peak increased with increasing reaction time and temperature, which signified the increase in the degree of acetylation of the ACE. There were similar thermal decomposition behaviors before and after esterification reaction until $800^{\circ}C$ so that the reaction occurred without significant structural changes of cellulose backbones. The solubility parameter of the ACE had a range of 18.5~26.4, and its viscosity and turbidity were controlled according to the solubility parameter of organic solvents. The ACE showed the hydrophilicity because the contact angle of the ACE was higher than the cellulose ether. These results confirmed that the ACE had the hydrophobicity and hydrophilicity due to the ether which was glucosidic bonding between the glucose units and un-reacted hydroxyl functional groups in the ACE.

Characterization of CO2 Gasification of 17 Coals With Regard to Coal Rank (다양한 등급의 17종 석탄의 CO2 가스화 반응특성 연구)

  • Kim, Soohyun;Yoo, Jiho;Chun, Donghyuk;Lee, Sihyun;Rhee, Young Woo
    • Clean Technology
    • /
    • v.19 no.3
    • /
    • pp.333-341
    • /
    • 2013
  • This paper presents results on $CO_2$ gasification of 17 raw coals containing a wide range of volatile matter (21-57 wt%). The gasification is performed using a TGA under $CO_2$ and also under $N_2$ atmosphere. An amount of weight loss with increasing temperature is proportional to that of volatile matter in a coal under $N_2$ atmosphere. Reactivity of $CO_2$ gasification also increases with a content of volatile matter. However, the correlation is a little scattered. Oxygenated functional groups in a coal are generally reactive and therefore, an increase in O/C ratio leads to enhanced reactivity. However, $CO_2$ reactivity is affected by neither H/C ratio nor a content of ashes that possibly activate the gasification reaction. These findings are also applicable to steam coal gasification and the reactivity series are confirmed in the test at a fixed bed reactor.

Isolation and Characterization of a Bacteriocin-Producing Lactobacillus sakei B16 from Kimchi (김치에서 박테리오신을 생산하는 Lactobacillus sakei B16의 분리 및 특성 분석)

  • Ahn, Ji-Eun;Kim, Jin-Kyoung;Lee, Hyeong-Rho;Eom, Hyun-Ju;Han, Nam-Soo
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.41 no.5
    • /
    • pp.721-726
    • /
    • 2012
  • Lactic acid bacteria (LAB) are able to secrete antimicrobial peptides called bacteriocins, which inhibit other bacteria such as pathogenic microorganisms. Therefore, bacteriocin-producing starters can be used as natural biopreservatives for various foods. The objective of this study was to screen and characterize bacteriocin-producing LAB from Kimchi and to investigate their applicability as a starter in Kimchi fermentation. To screen bacteriocin-producing LAB, gram-positive and gram-negative bacteria were used as indicators. To measure the antimicrobial activities of isolates, agar well diffusion assay method was used. According to the results, bacteriocin produced by $Lb.$ $sakei$ B16 showed antimicrobial activity against $Listeria$ $monocytogenes$ ATCC 19115, $Escherichia$ $coli$ KCTC 1467, and$Lactobacillus$ $plantarum$ KTCT 3104. Furthermore, bacteriocin was very stable after treatment with high temperature and high and low pH, but its effects were inhibited by treatment with proteolytic enzymes such as trypsin, proteinase K, and ${\alpha}$-chymotrypsin, revealing their bacteriocin-like protein- based structure. These results suggest that $Lb.$ $sakei$ B16 and its bacteriocin are good candidates as a functional probiotic and natural biopreservative, respectively, in fermented foods.

Development of Hyaluronic Acid-Functionalized Hydrogel Lens and Characterization of Physical Properties and Lysozyme Adsorption (Hyaluronic acid의 첨가방법에 따른 하이드로겔 콘택트렌즈의 물리적 특성과 lysozyme 흡착량 비교)

  • Lim, Hwa-lim;Kim, Ho-joong;Jun, Jin
    • Journal of Korean Ophthalmic Optics Society
    • /
    • v.20 no.3
    • /
    • pp.285-291
    • /
    • 2015
  • Purpose: The hydrogel lenses have been functionalized with HA(Hyaluronic Acid) using two different methods: construction of an IPN(Interpenetrating Polymer Networks) and formation of CCB(Chemical Covalent Bonding). The lysozyme adsorption and physical properties such as optical transmittance and water content of the hydrogel lenses have been investigated in order to determine whether method is suitable for the application potentials in contact lens industry. Methods: HA have been added to the hydrogel lenses prepared in the Lab using the two different method, e.g. IPN and CCB. The optical transmittance was measured in the wavelength range of 300~800 nm. The water content was measured by the gravimetric method using 0.9% NaCl saline solution. The amounts of adsorbed lysozyme on the contact lenses was analyzed by HPLC after incubation for 12h in artificial tears. Results: The water content of the HA added hydrogel contact lenses was increased, and the lens made by IPN method showed higher water content than the lens made by CCB method. The optical transmittance was over 90% both before and after addition of HA. Comparing the lysozyme adsorption reduction ratio, contact lens manufactured by IPN method was 60.0%, and the lens made by CCB method was 40.4%. Conclusions: CCB method is appropriate to distribute the functional material evenly throughout the lens, whereas IPN method is effective for the case of giving the functionality on the lens surface without phase separation.

Development of high tryptophan GM rice and its transcriptome analysis (고 함량 트립토판 생산 GM 벼 개발 및 전사체 분석)

  • Jung, Yu Jin;Nogoy, Franz Marielle;Cho, Yong-Gu;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
    • /
    • v.42 no.3
    • /
    • pp.186-195
    • /
    • 2015
  • Anthranilate synthase (AS) is a key enzyme in the biosynthesis of tryptophan (Trp), which is the precursor of bioactive metabolites like indole-3-acetic acid and other indole alkaloids. Alpha anthranilate synthase 2 (OsASA2) plays a critical role in the feedback inhibition of tryptophan biosynthesis. In this study, two vectors with single (F124V) and double (S126F/L530D) point mutations of the OsASA2 gene for feedback-insensitive ${\alpha}$ subunit of rice anthranilate synthase were constructed and transformed into wildtype Dongjinbyeo by Agrobacterium-mediated transformation. Transgenic single and double mutant lines were selected as a single copy using TaqMan PCR utilized nos gene probe. To select intergenic lines, the flanking sequence of RB or LB was digested with a BfaI enzyme. Four intergenic lines were selected using a flanking sequence tagged (FST) analysis. Expression in rice (Oryza sativa L.) of the transgenes resulted in the accumulation of tryptophan (Trp), indole-3-acetonitrile (IAN), and indole-3-acetic acid (IAA) in leaves and tryptophan content as a free amino acid in seeds also increased up to 30 times relative to the wildtype. Two homozygous event lines, S-TG1 and D-TG1, were selected for characterization of agronomic traits and metabolite profiling of seeds. Differentially expressed genes (DEGs), related to ion transfer and nutrient supply, were upregulated and DEGs related to co-enzymes that work as functional genes were down regulated. These results suggest that two homozygous event lines may prove effective for the breeding of crops with an increased level of free tryptophan content.