• Title/Summary/Keyword: galE

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Zygotic Expression of c-myc Gene in Mouse Early Embryos: Functional Role of c-myc Promoter (생쥐 초기배아에서 c-myc Proto-Oncogene Promoter의 기능적 활성화)

  • Park, Ki-Soo;Kang, Hae-Mook;Shim, Chan-seob;Sun, Woong;Kim, Jae-man;Lee, Young-Ki;Kim, Kyung-jin
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.550-556
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    • 1995
  • The c-myc proto-oncogene is Involved In the control of normal cell proliferation and differentiation of many cell lineages. Although it has heen suggested that c-myc may play an important role in the mammalian early development, it Is unclear whether the embryonic c-myc mRNA is originated from zygotic gene expression or stored maternal message. Thus, we have construded expression vectors, In which the 5, flanking sequences including c-myc promoter region and a large non-coding exon I are fused 'sith E. coli lacZ gene that encedes $\beta$-galactosldase as a reporter. As c-myc exon I contains a modulatory sequence, we designed t, vo types of vectors (pcmyc.Gall and pcmyc-Ga12) to examine the role of exon I in c-myc expression. The former contains the complete exon I and the later has a deletion in 40 bp of modulator sequence located In the exon I of c-myc These vectors were microInjected into fertilized one-cell embryos and $\beta$-galactosidase activity was examined by X-gal staining during early embryogenesis. $\beta$-galactosidase activity derived from c-myc promoter was decreased at two-cell stage. The expression level directed by pcmyc- Ga12 was similar to that of pcmyc-Gal1, indicating that the medulatory sequence in exon I may not be Involved at least In the regulation of embryonic c-myc expression. In summary, the present study indicates that the c-myc promoter is functional at the early stage embryo, and the regulation of c-myc expression is under the control of "zygotic" clock of preimplantation mouse embryos.e embryos.

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Change in Proteomic Profiles of Genetically Modified 1,3-Propanediol-Producing Recombinant E. coli

  • Jin, Li-Hua;Lee, Jung-Heon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1439-1444
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    • 2008
  • The recombinant E. coli $\Delta$6 mutant (galR, glpK, gldA, IdhA, lacI, tpiA) was used to produce 1,3-propanediol (PD) from glucose. The 1,3-PD production increased with feedback control of the glucose concentration using fed-batch fermentation. The maximum 1,3-PD concentration produced was 43 g/l after 60 h of fermentation. Glycerol production was minimized when controlling the glucose concentration at less than 1 g/l. The expression levels of seven enzymes related to the 1,3-PD production metabolism were compared during the cell growth phase and 1,3-PD production phase, and their expression levels all increased during 1,3-PD production, with the exception of alcohol dehydrogenase.

Molecular Cloning of $\beta$-Galactosidase from Bacillus subtilis HP-4

  • Kim, Jeong-Ho;Lee, Jae-Chang;Huh, Jeong-Won;Chung, Ki-Chul
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.227-231
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    • 1991
  • A gene coding for a $\beta$-galactosidase of Bacillus subtilis HP-4 was cloned in E. coli JM109 by inserting HindIII digested fragment of B. subtilis HP-4 chromosomal DNA into the site of pBR322 and selecting recombinant transformant showing blue color on X-gal plate. The recombinant plasmid, named pBG109, was found to contain the 1.4 Kbp HindIII fragment originated from B. subtilis HP-4 chromosomal DNA by Southern hybridization. The cloned gene was stably maintained and expressed in E. coli JM109 and the pBG109 encoded $\beta$-galactosidase had the same enzymatic properties as those of $\beta$-galactosidase produced by B. subtilis HP-4.

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MINIMAL DEL PEZZO SURFACES OF DEGREE 2 OVER FINITE FIELDS

  • Trepalin, Andrey
    • Bulletin of the Korean Mathematical Society
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    • v.54 no.5
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    • pp.1779-1801
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    • 2017
  • Let X be a minimal del Pezzo surface of degree 2 over a finite field ${\mathbb{F}}_q$. The image ${\Gamma}$ of the Galois group Gal(${\bar{\mathbb{F}}}_q/{\mathbb{F}}_q$) in the group Aut($Pic({\bar{X}})$) is a cyclic subgroup of the Weyl group W($E_7$). There are 60 conjugacy classes of cyclic subgroups in W($E_7$) and 18 of them correspond to minimal del Pezzo surfaces. In this paper we study which possibilities of these subgroups for minimal del Pezzo surfaces of degree 2 can be achieved for given q.

Metabolic Engineering of Escherichia coli for the Biological Synthesis of 7-O-Xylosyl Naringenin

  • Simkhada, Dinesh;Kim, EuiMin;Lee, Hei Chan;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.28 no.4
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    • pp.397-401
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    • 2009
  • Flavonoids are a group of polyphenolic compounds that have been recognized as important due to their physiological and pharmacological roles and their health benefits. Glycosylation of flavonoids has a wide range of effects on flavonoid solubility, stability, and bioavailability. We previously generated the E. coli BL21 (DE3) ${\Delta}pgi$ host by deleting the glucose-phosphate isomerase (Pgi) gene in E. coli BL21 (DE3). This host was further engineered for whole-cell biotransformation by integration of galU from E. coli K12, and expression of calS8 (UDP-glucose dehydrogenase) and calS9 (UDP-glucuronic acid decarboxylase) from Micromonospora echinospora spp. calichensis and arGt-4 (7-O-glycosyltransferase) from Arabidopsis thaliana to form E. coli (US89Gt-4), which is expected to produce glycosylated flavonoids. To test the designed system, the engineered host was fed with naringenin as a substrate, and naringenin 7-O-xyloside, a glycosylated naringenin product, was detected. Product was verified by HPLC-LC/MS and ESI-MS/MS analyses. The reconstructed host can be applied for the production of various classes of glycosylated flavonoids.

Genetic Characterization of the Escherichia coli O66 Antigen and Functional Identification of its wzy Gene

  • Cheng, Jiansong;Liu, Bin;Bastin David A.;Han, Weiqing;Wang, Lei;Feng Lu
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.69-74
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    • 2007
  • Escherichia coli is a clonal species, and occurs as both commensal and pathogenic strains, which are normally classified on the basis of their O, H, and K antigens. The O-antigen (O-specific polysaccharide), which consists of a series of oligosaccharide (O-unit) repeats, contributes major antigenic variability to the cell surface. The O-antigen gene cluster of E. coli O66 was sequenced in this study. The genes putatively responsible for the biosynthesis of dTDP-6-deoxy-L-talose and GDP-mannose, as well as those responsible for the transfer of sugars and for O-unit processing were identified based on their homology. The function of the wzy gene was confirmed by the results of a mutation test. Genes specific for E. coli O66 were identified via PCR screening against representatives of 186 E. coli and Shigella O type strains. The comparison of intergenic sequences located between galF and the O-antigen gene cluster in a range of E. coli and Shigella showed that this region may perform an important function in the homologous recombination of the O-antigen gene clusters.

Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection (외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.167-176
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    • 1997
  • This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

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Construction of a Lactococcal Shuttle/Expression Vector Containing a $\beta$-Galactosidase Gene as a Screening Marker (선별마커로써 $\beta$-Galactosidase 유전자를 포함한 Lactococcus용 셔틀/발현 벡터 제조)

  • Han Tae Un;Jeong Do-Won;Cho San Ho;Lee Jong-Hoon;Chung Dae Kyun;Lee Hyong Joo
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.241-247
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    • 2005
  • A new lactococcal shuttle/expression vector for lactococci, pWgal13T, was constructed using a $\beta$-galactosi-dase gene (lacZ) from Lacfococcus lactis ssp. lactis ATCC 7962 as a screening marker. The pWgal 13T was introduced into Escherichia coli DH5a and L. lactis MG1363, and was easily detected by the formation of blue colonies on a medium containing X-gal without any false transformants. Also, the quantitatively lacZ activity of pWgal13T was measured in L. lactis ssp. cremoris MG1363, and was found to be four times higher than that of L. lactis ssp. lactis ATCC7962 grown on a medium containing glucose, which shows that the lacZ gene of pWgal13T can be used for the efficient screening of L. lactis on general media. The pWgal13T was equipped with a lactococcal replicon of pWV01 from L. lactis Wg2, the new promoter P13C from L. lactis ssp. cremoris LM0230, multiple cloning sites, and a terminator for the expression of a relevant gene. The vee-tor pWgal13T was used for the expression of the EGFP gene in E. coli and L. lactis. These results show that the lactococcal expression/shuttle vector constructed in the present study can be used for the production of foreign proteins in E. coli and L. lactis.

Expression of Human Lactoferrin Gene and Secretion in Saccharomyces diastaticus YIY345 (효모 Saccharomyces diastaticus YIY 345에서의 Human Lactoferrin 유전자 발현 및 분비)

  • Joo, Yun Jung;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.23 no.1
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    • pp.80-89
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    • 1996
  • The expression and secretion of human lactoferrin (hLf) in Sacclnromyces diastaticus were performed. 1. For the secretion of hLf in yeast, recombinant plasmid pYEGLf was constructed using promoter, secretion signal sequence of glucoamylase I gene (STA1) and transcriptional terminator of GAL7 gene. 2. Each correct recombinant plasmid was selected by mini-preparation of plasmid DNA from E coli transformant and restriction enzyme digestion analysis. The selected plasmids, pYEGLf, were transformed into S. diastaticus YIY345 as a expression host, respectively. 3. Western blot analysis using rabbit anti-hLf was carried out to identify expressed hLf. Positive signals were shown in culture supernatant of pYEGLf transformant. 4. About $100{\mu}g-1mg$ of concentrated culture supernatant of positive clone were loaded on paper disc and tested for the antimicrobial activity against E coli. However, no activity was observed. We concluded that this fact results from low concentration of hLf secreted from yeast, compared with the fact that MIC of hLf is as high as $3mg/m{\ell}$. Therefore, the purification of secreted hLf may be require to investigate the antimicrobial activity. From this study, the feasibility of low-cost production of sufficient quantities of human lactofferin for nutritional and therapeutical applications were suggested.

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growth of Cadmium Sulfide (CdS) Thin Film by Solution Growth Technique and Study of Quantum Size Effects (용액성장법에 의한 Cadmium Sulfide(CdS) 박막 성장 및 양자 사이즈 효과에 관한 연구)

  • 임상철
    • Journal of the Microelectronics and Packaging Society
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    • v.4 no.1
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    • pp.1-12
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    • 1997
  • 본 연구에서는 용액성장법에 의해 양자 입자로 구성된 CdS 박막을 슬라이드 유리기 판위에 성장시키고 이들의 구조적 광학적 특성에 대하여 연구하였고 이들 결과를 토대로 용 액성장법으로 성장된 CdS 박막의 양자 사이즈 효과에 대하여 연구하였다. 성장시간은 1, 3, 10, 20분이었고 성장온도는 75$^{\circ}C$였다. X-선 회절 분석결과 본 연구에서 합성된 CdS 박막은 hexagonal상의 결정구조를 갖는 것으로 나타났고 성장시간에 따라 막의 투께는 61~195nm, 입자사이즈는 8.5~22.5nm로 나타났다. 광에너지 변화에 따른 투과도 측정결과 본 연구의 CdS 시료는 성장시간에 따라 에너지 밴드갭이 2.43~2.51 eV로 나타나서 벌크 CdS의 에너 지 밴드갭인 2.42 ev보다 높은 에너지 밴드갭을 갖게 되어 양자 사이즈 효과에 의한 blue shift 현상이 용액성장법에 의해 합성된 CdS 시료에도 존재한다는 것이 밝혀졌다 그리고 이 같은 용액성장법으로 성장된 CdS에 대해 최초로 수행된 Raman 산란 실험결과 이성장방법 으로 성장된 CdS에는 1TO, E2, 1LO 포논 모드가 존재함을 알수 있었고 또한 입자 사이즈 감소에 의한 1LO포논 모드의저주파수 shift 현상 즉 포논 모드의 softening 현상이 있음이 밝혀졌고 softening은 최대6.2%까지 발생하였다. 이와같은 높은 softening은 본연구의 CdS 박막 내 양자 입자의 입도가 작은것에 기인하는 것으로 밝혀졌다. 또한 본 CdS 시료의 양 자 사이즈 효과의 결과로 1TO 포논도 나타났는데 이 1TO 포논과 E2 포논의 Raman shift 는 성장시간 즉 막의 두께와는 무관한 것으로 나타났다.행렬모형(二重比例行列模型)을 이용하여, 산업구조의 변화로 인한 직업별 인력수요 변화가 충분히 고려되도록 하였다. 전망의 결과에 따르면 향후 우리 경제는 지식기반경제(knowledge-based economy)로 이행하고 있다고 볼 수 있다. 우선 산업구조면에서 지식집약적산업으로의 구조조정이 일어나게 되고 이에 따라 산업별 취업구조에서도 고기술산업의 취업준비중이 급속히 증가하게 된다. 직업별 취업분포에 있어서도 전문기술직 행정관리직 등의 고숙련 사무직의 비중은 크게 증가하는 반면 생산관련직과 농림어업직의 비중은 감소하게 된다. 이처럼 경제가 지식집약화되어 감에 따라 고학력자에 대한 수요는 지속적으로 증가하지만 현재 적절한 인력양성과 공급이 이루어지지 않고 있어 향후 기술이나 기능에 따른 수급부일정(需給不一政)(skill mismatch)현상이 매우 심해질 것으로 보인다. 따라서 앞으로의 인력정책에서 가장 주안점을 두어야 할 부분은 첨단기술산업과 관련된 인력의 양성에 있다고 하겠다.2시간까지 LPDG용액은 MEC용액보다 비교적 나은 회복을 보였고 재관류 3일과 7일의 폐기능 평가에서 두 용액 모두에서 폐기능의 점차적 소실을 보였으며 이는 병리조직검사에서 보듯이 폐혐에 의한 외적인 요소라고 생각되며 따라서 LPDG용액은 허혈재관류손상 방지 및 급성폐렴 등 염증을 잘 관리한다면 20시간 이상 LPDG용액의 안전한 폐보존의 가능성 을 얻을 수 있었다.ic 형태로 외래유전자가 발현되었지만 대조구에서 87.0% (26/30개) 배반포기가 $\beta$-Gal 활력을 보인 반면, G418 처리구에서는 모든 배반포기가 $\beta$-Gal 활력을 보였다 (P<0.05). 그러나 대조구 및 G418 처리구의 ICM