• Title/Summary/Keyword: lac I

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Post-pillars design for safe exploitation at Trepça hard rock mine (Kosovo) based on numerical modeling

  • Ibishi, Gzim;Genis, Melih;Yavuz, Mahmut
    • Geomechanics and Engineering
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    • v.28 no.5
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    • pp.463-475
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    • 2022
  • In the mine exploitation stage; one of the critical issues is the stability assessment of post-pillars. The instability of post-pillars leads to serious safety hazards in mining operations. The focus of this study is to assess the stability of post-pillars in the 130# stope in the central ore body at Trepça hard rock mine by employing both conventional (i.e., critical span curve) and numerical methods (i.e., FLAC3D). Moreover, a new numerical based index (i.e., Pillar Yield Ratio-PYR) was proposed. The aim of PYR index is to determine a border line between stable, potentially unstable, and failure state of post-pillars at a specific mine site. The critical value of pillar width to height ratio is 2.5 for deep production stopes (e.g., > 800 m). Results showed that pillar size, mining height and mining depth significantly have affected the post-pillar stability. The reliability of numerical based index (i.e., PYR) is verified based on empirical underground pillar stability graph developed by Lunder, 1994. The proposed pillar yield ratio index and pillar stability graph can be used as a design tool in new mining areas at Trepça hard rock mine and for other situations with similar geotechnical conditions.

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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cdma2000 Physical Layer: An overview

  • Willenegger, Serge
    • Journal of Communications and Networks
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    • v.2 no.1
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    • pp.5-17
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    • 2000
  • cdma2000 offers several enhancement as compared to TIA/EIA-95, although it remains fully compatible with TIA/EIA-95 systems and allows for a smooth migration from one to the other-Major new capability include:1)connectivity to GSM-MAP in addition to IP and IS-41 networks; 2) new layering with new LAC and MAC architectures for improved service multiplexing and QoS management and efficient use of radio resource ;3) new bands and band widths of operation in support of various operator need and constraints, as well as desire for a smooth and progressive migration to cdma 2000; and 4) flexible channel structure in support of multiple services with various QoS and variable transmission rates at up to 1 Mbps per channel and 2 Mbps per user. Given the phenomenal success of wireless services and desire for higher rate wireless services. improved spectrum efficiency was a major design goal in the elaboration of cdma2000. Major capacity enhancing features include; 1) turbo coding for data transmission: 2)fast forward link power control :3) forward link transmit diversity; 4) support of directive antenna transmission techniques; 5) coherent reverse link structure; and 6) enhanced access channel operation. As users increasingly rely on their cell phone at work and at home for voice and data exchange, the stand-by time and operation-time are essential parameters that can influence customer's satisfaction and service utilization. Another major goal of cdma2000 was therefore to enable manufacturers to further optimize power utilization in the terminal. Major battery life enhancing features include; 1) improved reverse link performance (i.e., reduced transmit power per information bit; 2) new common channel structure and operation ;3) quick paging channel operation; 4) reverse link gated transmission ; and 5) new MAC stated for efficient and ubiquitous idle time idle time operation. this article provides additional details on those enhancements. The intent is not to duplicate the detailed cdma2000 radio access network specification, but rather to provide some background on the new features of cdma2000 and on the qualitative improvements as compared to the TIA/EIA-95 based systems. The article is focused on the physical layer structure and associated procedures. It therefore does not cover the MAC, LAC, radio resource management [1], or any other signaling protocols in any detail. We assume some familiarity with the basic CDMA concepts used in TIA/EIA-95.

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Characterization of gp64 Gene of Bombyx mori Nucleopolyhedrovirus and Development of a Transient Expression Vector (누에 핵다각체병 바이러스 헤 gp64 유전자의 특성조사 및 transient 발현 벡터 개발)

  • 김미향;최재영;우수동;이해광;제연호
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.18-24
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    • 2001
  • Expression of the baculovirus major envelope glycoprotein gene(gp64) is regulated by transcription from botha early and late promoters. To develop a transient expression vector under the control of gp64 gene promoter, the gp64 gene of Bombyx mori nucleopolyhedrovirus-K1(BmNPV-K1) was characterized. The gp64 gene was local-ized at EcoR I-Pst I 7.38-kb fragment of the BmNPV-K1 genome. The EcorR 1-Pst I 7.38-kb fragment was cloned and the nucleotide sequence of 2,277 bases including the coding region of gp64 gene was determined. Based on these results, transient expression vector using gp64 gene promoter was constructed and named as pBm64. E.coli lacZ gene was introduced onto pBm64 as a reporter gene and expressed transiently in B. mori 5(Bm 5) cells. The expression vector transfected into the cells was maintained stably for 1 to 5 days. In order to confirm the expression of the reporter gene by gp64 promoter, recombinant virus was constructed. The recombinant virus has two independent transcription units in opposite orientations with two promoters; gp64 and polyhedrin gene promoters each initiating transcription of $\beta$-galactosidase and polyhedrin, respectively. Polyhedra formation and expression of $\beta$-galactosidase in Bm5 cells infected with the recombinant virus were observed with phase contrast microscope and in situ staining.

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Glutamic Acid Rich Helix II Domain of the HIV-1 Vpu has Transactivation Potential in Yeast

  • Hong, Seung-Keun;Bae, Yong-Soo;Kim, Jung-Woo
    • BMB Reports
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    • v.32 no.4
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    • pp.405-408
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    • 1999
  • The transactivation potential of HIV-1 Vpu was identified from the yeast two-hybrid screening process. The helix II domain of HIV-1 Vpu protein and mutant Vpu protein lacking the transmembrane domain exhibited transactivation of the LacZ and Leu2 reporter genes carrying LexA upstream activating sequences, but full-length HIV-1 Vpu and the helix I domain of HIV-1 Vpu did not. The helix II domain of HIV-1 Vpu consists of a number of acidic amino acids, and is especially rich in glutamic acid, a characteristic of many transcription factors. This result suggests that protein-protein interaction may occur through the acidic helix II domain of HIV-1 Vpu.

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An analysis of the arm-type site binding domain of bacteriophage .lambda. integrase

  • Cho, Eun-Hee
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.165-170
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    • 1995
  • The 356 amino acid long lambda integrase protein of bacteriophage .lambda. constains two autonomous DNA binding domains with distinct sequence specificities. The amino terminal domain of integrase is implicated to bind to the arm-type sequences and the carboxyl domain interacts with the coretype sequencess. As a first step to understand the molecular mechanism of the integrase-DNA interaction at the arm-type site, the int(am)94 gene carrying an amber mutation at the 94th codon of the int was cloned under the control of the P$\_$tac/ promoter and the lacI$\_$q/ gene. The Int(am)94 mutant protein of amino terminal 93 amino acid residues can be produced at high level from a suppressor free strain harboring the plasmid pInt(am)94. The arm-type binding activity of Int(am)94 were measured in vivo and in vitro. A comparison of the arm-type binding properties of the wild-type integrase and the truncated Int(am)94 mutant indicated that the truncated fragment containing 93 amino acid residues carry all the determinants for DNA binding at the arm-type sites.

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A Modified PCR-Directed Gene Replacements Method Using $lambda$-Red Recombination Functions in Escherichia coli

  • KIM SANG-YOON;CHO JAE-YONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1346-1352
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    • 2005
  • We have developed a modified gene replacement method using PCR products containing short homologous sequences of 40- to 50-nt. The method required $\lambda$-Red recombination functions provided under the control of a temperature-sensitive CI857 repressor expressed from the $P_{lac}$ promoter in the presence of IPTG on an easily curable helper plasmid. The method promoted the targeted gene replacements in the Escherichia coli chromosome after shifting cultures of the recombinogenic host, which carries the helper plasmid, to $42^{\circ}C$ for 15 min. Since this method employs $\lambda$-Red recombination functions expressed from the easily curable helper plasmid, multiple rounds of gene replacements in the E. coli chromosome would be possible. The procedures described herein are expected to be widely used for metabolic engineering of E. coli and other bacteria.

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.

Mutation Spectrum of 1, 2-Dibromo-3-chloropropane, an Endocrine Disruptor, in the lacI Transgenic Big $\textrm{Blue}^{(R)}$ Rat2 Fibroblast Cell Line.

  • Kim, Youn-Jung;Chai, Young-Gyu;Lee, Kilchul;Kyounghee Oh;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2001.05a
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    • pp.129-129
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    • 2001
  • 1,2-Dibromo-3-chloropropane (DBCP), a soil fumigant against nematodes, is a genotoxic carcinogen and also is classified by World Wildlife Fund as endocrine disruptors. DBCP has been extensively studied on genotoxicity, carcinogenicity, and damage in male reproductive-related organs. However, information on precise mechanism of mutagenesis and carcinogenesis of DBCP is yet unknown. (omitted)

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Molecular cloning of phospho-$\beta$-galactosidase gene of lactobacillus casei in escherichia coli (Lactobacillus casei의 phospho-$\beta$-galactosidase 유전자의 대장균내 분자클로닝)

  • 문경희;박정희;최순영;이유미;김태한;김연수;민경희
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.188-193
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    • 1989
  • Gene for lactose catabolism in Lactobacillus casei SW-M1 was encoded by a 60Kb metabolic plasmid. A derivative of only 10kb, pPlac 15 of recombinant plasmid, was constructed by introducing into pBR322 and was cloned into E. coli using restriction endonuclease Pst I. A 10kb insery DNA in plasmid pBR322 was identified as a gene encoded phospho-$\beta$-galactosidase by the determination of enzyme activity. Phospho-$\beta$-galactosidase was apparently expressed in E. coli. The enzyme activities of cell-free extract from transformant E. coli HB101 carrying pPLac 15 DNA were not different from that of L. casei as a donor strain on the basis of enzyme properites. However, specific activity of phospho-$\beta$-galactosidase in the cloned strain with Lac $Y^{-}$ phenotype of E. coli HB101 was lower than that in L. casei strain.

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