• 제목/요약/키워드: galE

검색결과 143건 처리시간 0.028초

동해 지역의 프리에어 이상으로부터 완전부우게 이상의 계산 (Computation of Complete Bouguer Anomalies from Free-air Anomalies in East Sea)

  • 윤홍식;이동하;김용현
    • 한국측량학회지
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    • 제28권3호
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    • pp.317-328
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    • 2010
  • 본 연구는 Sandwell 및 DNSC08 해상중력모델로부터 구한 프리에어 이상으로부터 동해 지역의 완전부우게 이상을 구한 결과를 설명한 것이다. 완전부우게 보정은 부우게 보정(Bullard A), 곡률보정(Bullard B)과 지형보정(Bullard C)의 세 부분으로 구성된다. 각 보정량의 계산을 위하여 전지구 기복모델인 ETOPO1을 통해 1분 간격의 표고데이터(지형 및 수심)를 취득하여 이용하였으며, 지형(해수)의 밀도로는 $2,670kg/m^3$($1,030kg/m^3$)의 수치를 일괄적으로 적용하였다. DNSC08 모델을 이용하여 계산된 완전부우게 이상은 동해 지역에 대하여 약 -34.390~267.925mGal의 분포를 나타내었으며, Sandwell 모델의 경우 -32.446~266.967mGal의 분포를 나타내었다. 또한, 두 모델 간 완전부우게 이상의 차이는 평균 $0.036{\pm}2.373mGal$로 계산되었으며, 가장 큰 완전부우게 이상값은 가장 낮은 수심분포를 보이는 위도 $42{\sim}43^{\circ}N$ 및 경도 $137{\sim}139^{\circ}E$ 사이의 지역에서 나타났다. 이러한 수치를 통해 DNSC08과 Sandwell 모델의 중력분포가 동해 지역에 대해 매우 유사한 것을 알 수 있었으며, 위성기반의 해상중력모델이 동해 지역의 지구물리학적, 지질학적, 측지학적 특성의 해석에 효율적으로 이용될 수 있다고 판단되었다.

미꾸라지, Misgurnus mizozepis에 외래 유전자 이식 I. lacZ의 reporter 유전자로서의 유용성 검토 (Transfer of Foreign Gene into Mud Loach, Misgurnus mizolepis I . Availability of the lacZ as a reporter gene for producing transgenic mud loach)

  • 김동수;남윤권
    • 한국양식학회지
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    • 제7권1호
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    • pp.41-54
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    • 1994
  • E. coli의 \beta-galactosidase$ 유전자를 미꾸라지 수정난에 미세현미 주입하고 이를 분석함으로써 미꾸라지에 외래 유전자 이식을 위한 reporter 유전자로서의 유용성을 검토하였다 X-gal 염객분석, 4-methylumbelliferyl-$\beta$-D-galactoside (MUG) 분석을 수행한 결과 유전자 이식 처리군 및 대조군에서 모두 \beta-galactosidase$의 활성이 관찰되었으며 PCR, dot blot 및 southern blot분석결과 역시 유전자 이식 처리군과 대조군에서 모두 유사한 양상을 나타내었다. 처리군 및 대조군의 PCR product의 염기서열은 E. coli의 \beta-galactosidase$ 유전자와 매우 높은 homology를 갖고 있었으며 pH에 따른 X-gal 염색 분석을 수행한 결과 미꾸라지에 관찰되는 본 효소는 pH 4.5에서 가장 높은 활성을 나타내었다. 따라서 앞으로 미꾸라지를 대상으로 한 외래 유전자 이식시 E. coli의 \beta-galactosidase$ 유전자의 reporter 유전자로서의 사용은 신중한 재검토가 이루어져야만 할 것으로 판단된다.

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감마선 조사된 난백 Albumin(Gal d 1)의 알러지성 및 항원성의 변화 (The Changes of Allergenic and Antigenic Properties of Egg White Albumin (Gal d 1) by Gamma Irradiation)

  • 이주운;육홍선;조경환;이수영;변명우
    • 한국식품영양과학회지
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    • 제30권3호
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    • pp.500-504
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    • 2001
  • Gamma irradiation was applied to reduce egg allergy. Ovalbumin (OVA), an egg white protein, was used as model allergen and was gamma-irradiated at 3, 5, or 10 kGy in an aqueous state (2.0 mg/mL). The changes in allergenic and antigenic properties of OVA resulted from gamma irradiation were monitored by ELISA with serum from egg-hypersensitive patients (H-IgE), and mouse monoclonal IgG (M-IgG) or rabbit polyclonal IgG (R-IgG). The binding ability of H-IgE to irradiated OVA was dose-dependently reduced. However, IgGs from animal did better recognize 3 or 5 kGy-irradiated OVA. In the evaluation of immune reactivity using blind test, the reactivity of H-IgE rapidly decreased depending upon the irradiation dose. However, the reactivities of M-IgG and R-IgG was higher at 5 and 3 kGy-irradiated OVA than non-irradiated control. The results provide a new possibility to use irradiation process for reducing the allergenicity of egg white.

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Yeast 2 $\mu$m 플라스미드 유래 FLP recombinase 유전자의 곤충 배양세포내 발현 (Expression of the FLP recombinase of the 2 $\mu$m plasmid of yeast in the cultured cells of Bombyx mori using a transient expression vector)

  • 강석우;윤은영
    • 한국잠사곤충학회지
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    • 제39권1호
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    • pp.36-43
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    • 1997
  • In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

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동해 지역의 완전부우게 이상 계산 (Computation of Complete Bouguer Anomalies in East Sea)

  • 김용현;윤홍식;이동하;황학
    • 한국측량학회:학술대회논문집
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    • 한국측량학회 2010년 춘계학술발표회 논문집
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    • pp.165-168
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    • 2010
  • This paper describes the results of complete Bouguer anomalies computed from the Free-air anomalies that derived from Sandwell and DNSC08 mairne gravity models. Complete bouguer corrections consist of three parts: the bouguer correction (Bullard A), the curvature correction (Bullard B) and the terrain correction (Bullard C). These all corrections have been computed over the East Sea on a $1'{\times}1'$ elevation data (topography and bathymetry) derived from ETOPO1 global relief model. In addition, a constant topographic (sea-water) density of $2,670kg/m^3$ ($1,030kg/m^3$) has been used for all correction terms. The distribution of complete bouguer anomalies computed from DNSC08 are -34.390 ~ 267.925 mGal, and those from Sandwell are -32.446 ~ 266.967 mGal in East Sea. The mean and RMSE value of the difference between DNSC08 and Sandwell is $0.036{\pm}2.373$ mGal. The highest value of complete bouguer anomaly are found around the region of $42{\sim}43^{\circ}N$ and $137{\sim}139^{\circ}E$ (has the lowest bathymetry) in both models. Theses values show that the gravity distribution of both models, DNSC08 and Sandwell, are very similar. They indicate that satellite-based marine gravity model can be effectively used to analyze the geophysical, geological and geodetic characteristics in East Sea.

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Effective Blocking of Microbial Transcriptional Initiation by dCas9-NG-Mediated CRISPR Interference

  • Kim, Bumjoon;Kim, Hyun Ju;Lee, Sang Jun
    • Journal of Microbiology and Biotechnology
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    • 제30권12호
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    • pp.1919-1926
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    • 2020
  • CRISPR interference (CRISPRi) has been developed as a transcriptional control tool by inactivating the DNA cleavage ability of Cas9 nucleases to produce dCas9 (deactivated Cas9), and leaving dCas9 the ability to specifically bind to the target DNA sequence. CRISPR/Cas9 technology has limitations in designing target-specific single-guide RNA (sgRNA) due to the dependence of protospacer adjacent motif (PAM) (5'-NGG) for binding target DNAs. Reportedly, Cas9-NG recognizing 5'-NG as the PAM sequence has been constructed by removing the dependence on the last base G of PAM through protein engineering of Cas9. In this study, a dCas9-NG protein was engineered by introducing two active site mutations in Cas9-NG, and its ability to regulate transcription was evaluated in the gal promoter in E. coli. Analysis of cell growth rate, D-galactose consumption rate, and gal transcripts confirmed that dCas9-NG can completely repress the promoter by recognizing DNA targets with PAM of 5'-NGG, NGA, NGC, NGT, and NAG. Our study showed possible PAM sequences for dCas9-NG and provided information on target-specific sgRNA design for regulation of both gene expression and cellular metabolism.

Amoebic PI3K and PKC Is Required for Jurkat T Cell Death Induced by Entamoeba histolytica

  • Lee, Young Ah;Kim, Kyeong Ah;Min, Arim;Shin, Myeong Heon
    • Parasites, Hosts and Diseases
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    • 제52권4호
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    • pp.355-365
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    • 2014
  • The enteric protozoan parasite Entamoeba histolytica is the causative agent of human amebiasis. During infection, adherence of E. histolytica through Gal/GalNAc lectin on the surface of the amoeba can induce caspase-3-dependent or -independent host cell death. Phosphorylinositol 3-kinase (PI3K) and protein kinase C (PKC) in E. histolytica play an important function in the adhesion, killing, or phagocytosis of target cells. In this study, we examined the role of amoebic PI3K and PKC in amoeba-induced apoptotic cell death in Jurkat T cells. When Jurkat T cells were incubated with E. histolytica trophozoites, phosphatidylserine (PS) externalization and DNA fragmentation in Jurkat cells were markedly increased compared to those of cells incubated with medium alone. However, when amoebae were pretreated with a PI3K inhibitor, wortmannin before being incubated with E. histolytica, E. histolytica-induced PS externalization and DNA fragmentation in Jurkat cells were significantly reduced compared to results for amoebae pretreated with DMSO. In addition, pretreatment of amoebae with a PKC inhibitor, staurosporine strongly inhibited Jurkat T cell death. However, E. histolytica-induced cleavage of caspase-3, -6, and -7 were not inhibited by pretreatment of amoebae with wortmannin or staurosporin. In addition, we found that amoebic PI3K and PKC have an important role on amoeba adhesion to host compartment. These results suggest that amebic PI3K and PKC activation may play an important role in caspase-independent cell death in Entamoeba-induced apoptosis.

Prevalence of virulence and cytolethal distending toxin (CDT) genes in thermophilic Campylobacter spp. from dogs and humans in Gyeongnam and Busan, Korea

  • Cho, Hyun-Ho;Kim, Sang-Hyun;Min, Wongi;Ku, Bok-Kyung;Kim, Yong-Hwan
    • 대한수의학회지
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    • 제54권1호
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    • pp.39-48
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    • 2014
  • The prevalence of thermophilic Campylobacter (C.) spp. in stray, breeding, and household dogs was 25.2, 12.0, and 8.8%, respectively. C. jejuni and C. upsaliensis were the predominant Campylobacter spp. from household dogs. cdtA, cdtB, and cdtC were detected by PCR in all isolates. Despite the high cytolethal distending toxin (CDT) gene prevalence, only 26 (31%) C. jejuni strains and one (15.3%) C. coli strain showed evidence of CDT production in HEp-2 cell cytotoxicity assays. Virulence-associated genes detected in the C. jejuni and C. coli isolates were cadF, dnaJ, flaA, racR, ciaB, iamA, pldA, virB11, ceuE, and docC. cadF, dnaJ, flaA, and ceuE were found in all C. jejuni and C. coli isolates. When detecting Guillain-Barr$\acute{e}$ syndrome-associated genes (galE, cgtB, and wlaN), galE was identified in all isolates. However, cgtB and wlaN were more prevalent in C. jejuni isolates from humans than those from dogs. Adherence and invasion abilities of the C. jejuni and C. coli strains were tested in INT-407 cells. A considerable correlation (adjusted $R^2$= 0.678) existed between adherence and invasion activities of the Campylobacter spp. isolates.

Expression of $\beta$-Galactosidase Gene of Lactococcus lactis ssp. lactis ATCC 7962 in Lactococcus lactis ssp. lactis MG1363

  • Park, Rae-Jun;Lee, Jung-Min;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Jong-Hoon;Lee, Hyong-Joo;Kim, Jeong-Hwan
    • Preventive Nutrition and Food Science
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    • 제5권3호
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    • pp.153-159
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    • 2000
  • A 4.4 kb DNA fragment encompassing lacA (galactoside acetyltransferase) and lacZ($\beta$-galactosidase) genes from Lactococus lactis ssp. lactis ATCC 7962 (L. lactis 7962) was introduced ito a Lac strain, Lactococcus lactis ssp. lactis MG1363 (L. lactis MG1363) by using a lactococcal expression vector, pMG36e and expression level of lacZ was examined. Growth rates and $\beta$-galactosidase ($\beta$-gal) activities of MG1363 cells carrying recombinant plasmid, pMLZ3, on M17 broth containing different carbon sources (1%, w/v) were examined. Contrary to the expectations, MG1363 [pMLZ3] grown on lactose showed the lowest enzyme activity (17 units) and cells grown on galactose had the highest $\beta$-gal activity (41 units). Cells grown on glucose had intermediate activity (33 units). These activities are about one tenth of the values observed in L. lactis 7962 where lacZ is present as a single-copy gene in the chromosome. When the cellular concentrations of lacZ transcript were examined using slot blot hybridization, it was found that MG1363[pMLZ3] produced sufficient amounts of transcript. These results indicate that either proteolytic degradation of $\beta$-gal or other regulatory mechanism prevent the translation or accumulation of $\beta$-gal in L. lactis MG1363 cells. In regard to regulation, the presence of the ccpA gene in L. lactis MG1363 was confirmed by Southern blot.

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Biophysical Feature, Crystallization and X-ray Crystallographic Studies of Toxascaris leonina Galectin

  • Sung, Min-Kyung;Jeong, Mi-Suk;Lee, Woo-Chul;Song, Jeong-Hyun;Kim, Hye-Yeon;Cho, Min-Kyoung;Yu, Hak-Sun;Jang, Se-Bok
    • Bulletin of the Korean Chemical Society
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    • 제33권1호
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    • pp.227-232
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    • 2012
  • Galectins are generally believed to be potential candidates for use in the development of novel antiinflammatory agents or as selective modulators of the immune response. In particular, galectin-9 exhibits some of the extracellular functions, including cell aggregation, adhesion, chemoattraction, activation, and apoptosis. Tl-galectin (Tl-gal, galectin-9 homologue gene) was isolated from an adult worm of the Toxascaris leonina. The full-length Tl-gal gene, which was incorporated into pET-28a, was overexpressed in E. coli and purified by nickel affinity and gel filtration chromatographies. The purified Tl-gal was crystallized using the hangingdrop vapor-diffusion method. The crystal belonged to the tetragonal space group $P4_1$, with unit-cell parameters of a = b = $75.7\AA$ and c = $248.4\AA$. The crystals were obtained at $20^{\circ}C$ and diffracted to a resolution of $3.0\AA$. The asymmetric unit contained four molecules of Tl-gal, which gave a crystal volume per protein mass (Vm) of $2.8\AA^3Da^{-1}$ and a solvent content of 54.1%.