• Title/Summary/Keyword: P.C strand

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Identification of Six Single-Strand Initiation (ssi) Signals for Priming of DNA Replication in Various Plasmids

  • Jeong, Jin-Yong;Seo, Hak-Soo;Kim, Ho-Yeon;Cho, Moo-Je;Bahk, Jeong-Dong
    • BMB Reports
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    • v.28 no.4
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    • pp.336-341
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    • 1995
  • Using a mutant M13 phage derivative lacking a great part of the complementary strand synthesis origin, we identified six single-strand initiation (ssi) signals for DNA replication in pACYC184, pLG214, pGKV21, and pDPT270 plasmids, and named them $ssiA_{YC}$, $ssiA_{LG}$, $ssiB_{LG}$, $ssiA_{KV}$, $ssiA_{PT}$, and $ssiB_{PT}$, respectively. Two of them were from pDPT270, one from downstream the on of pACYC184, two from pLG214, one from upstream the plus origin of pGKV21. Introduction of these ssi signals into the deleted $ori_c$ site of a mutant filamentous M13 phage ($M13{\Delta}lac182$) resulted in the restoration of growth activity of this phage. These ssi signals were classified into a number of groups on the basis of sequence similarity. $ssiA_{YC}$ and $ssiA_{LG}$ show extensive sequence homology to the n'-site (primosome assembly sites) of ColE1, whereas $ssiB_{PT}$ is homologous to the n'-site of ${\Phi}X174$. $ssiA_{PT}$ belongs to G4-type ssi signals which require only dnaG primase and SSB protein for the priming of replication. In addition, possible biological roles of these ssi signals are discussed.

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Molecular Cloning of H-Y Antigen Gene III. Construction of Mouse Testis cDNA Library and Screening of H-Y Ag Gene (H-Y 항원 유전자의 클로닝에 관한 연구 III. 생쥐정소 cDNA Library 구성과 유전자의 검색)

  • 이정렬;김창규;김종배
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.43-48
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    • 1993
  • These experiments were carried out to construct mouse testis cDNA library and to to seen H-Y Ag gene. Mouse testis was obtained from BALB/c inbreed mouse that was after-born 1 week. Isolation of mouse testis total RNA was carried out by guanidum/cesium choloride, poly(A+) mRNAs were purified by oligo d(T)-cellulose chromatography method. To investigate protein synthesis activity, in-vitro translation carried out by total RNA and poly(A+) mRNA. The products of in-vitro translation were identified in 12.5% PAGE. Single strand DNA and double strand DNA were synthesized from poly(A+) mRNA and purified using phenol/chloroform/isoamylalcohol. Synthesized cDNA was combined with cohesive Eco RI polylinker, its recombination efficiencies were identified by X-gal and IPTG. In the cDNA library, 1$\times$107 phagemids were screened with 32P labelled probe. Hybridization were carried on $65^{\circ}C$ for 16~20hours. And 1$\times$106 phagemids were screened with rabbit-anti-H-Y. In former, select 5 positive clones, and later, 1 positive clone. Its southern blot analysis showed various size of insert cDNA from 0.7kb to 3kb.

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Replication origin (ori) of R-plasmid pSBK203 Isolated from Staphylococcus aureus DHI (Staphylococcus aureus DH1에서 분리한 R-plasmid pSBK203상의 복제개시 부위 ori에 관한 연구)

  • Min, Kyung-Il;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.186-191
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    • 1994
  • The origin of the leading strand replication (ori) and of lagging strand replication (M-O) of R-plasmid pSBK203 was identified and its base sequence was determined. About 50 bp of ori sequence residues overlapped with the structural gene of rep. Sequence comparison reveals that pSBK-ori shares obvious identities with those of pT181 family and consists of two regions, one is conserved and the other is variable region. Of two palindrome sequence located one after another in upstream region of rep gene, palA' instead of palA which shares sequence homology with diverse family of plasmids such as pOX6, pC194, and pE194 seems to act as a signal for conversion of primarily replicated ssDNA to dsDNA (minus origin (M-O)).

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A Study on the Calculation of Load Resistance Factor of over Tension Anchors by Optimization Design (최적화 설계를 통한 과긴장 앵커의 하중-저항계수 산정 연구)

  • Soung-Kyu Lee;Yeong-Jin Lee;Yong-Jae Song;Tae-Jun Cho;Kang-Il Lee
    • Journal of the Korean Geosynthetics Society
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    • v.22 no.4
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    • pp.17-26
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    • 2023
  • To consider the risk of damage and fracture of P.C strands, the existing post-maintenance system alone has the limitations, hence it is necessary to quantitatively evaluate and predict the deterioration, durability and safety of facilities and establish a reasonable maintenance system considering the asset value of facilities. Therefore, it is worth considering a preventive maintenance plan that allows proactive measures to be taken before a major defect occurs in the temporary anchor. This study devised a preventive over tension method, reviewed its effectiveness through design and field tests, by calculating the resistance factors by performing a reliability-based optimization design. At this time, the over tension anchor method was evaluated using the ratio of the residual tension force after the fracture of P.C strands to the effective tension force before the fracture of P.C strand, followed by the resistance factor calculated by the optimal solution for each random variables using Excel solver and applying it to the limit state equations. As a result of the study, if the over tension ratio is 125% to 130%, the remaining strands showed a high resistance effect even after the fracture of P.C strand. As a result of the optimization design, it was found that it is appropriate to apply the load factor (γ) of 1.25, and the resistance factors of Φ1, Φ2, Φ3 as 0.7, 0.5, 0.6.

고체추진제의 Thermal Structure 분석에 관한 실험적 연구

  • 채재우;문종인
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 1996.05a
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    • pp.109-117
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    • 1996
  • Strand Burner 내에 설치되는 추진제에 10$\mu\textrm{m}$굵기의 Thermocouple을 설치하여 각 압력하에서의 연소속도 및 연소시의 Thermal Structure의 정보를 얻는 방법과 러시아의I.C.P에서 공동실험을 통하여 얻은 60%의 니트로글리세린과 40%의 니트로셀룰로오스를 혼합한 더블베이스 추진제 및 AP와 에폭시수지를 혼합한 간이 Composite형 추진제의 Thermal Structure를 조사한다.

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Detection of DNA Adduct Formed by Mitomycin C by $^{32}P$-Postlabelling ($^{32}P$-Postlabelling 방법을 이용한 미토마이신 C에 의하여 형성된 DNA adduct의 검출)

  • Jeong, Hye-Yun;Kim, Jae-Hyeon;Park, Chang-Won;Lee, Dong-Gwon
    • YAKHAK HOEJI
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    • v.40 no.4
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    • pp.442-448
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    • 1996
  • Mitomycin C(MMC) has been used as an anticancer drug and behaves as an alkylating agent forming covalent cross-link between complementary strands of double strand DNA. The purpose of this research was to determine number of DNA adducts, formed in vivo by Mitomycin C, in mouse organs. DNAs from liver, lung, brain and pancreas were isolated and used for $^{32}P$-postlabelling. The labeled nucleotides were separated by 2D-TLC and subjected to autoradiography. Numbers of MMC-DNA adducts were 9,9,5,4 in liver, pancreas, lung and brain, respectively.

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Effects of Rutaecarpine on Hydrogen Peroxide-Induced Apoptosis in Murine Hepa-1c1c7 Cells

  • Lee, Sung-Jin;Ahn, Hyun-Jin;Nam, Kung-Woo;Kim, Kyeong-Ho;Mar, Woong-Chon
    • Biomolecules & Therapeutics
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    • v.20 no.5
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    • pp.487-491
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    • 2012
  • The aim of this study was to investigate the inhibitory effects of rutaecarpine on DNA strand breaks and apoptosis induced by hydrogen peroxide ($H_2O_2$) in murine Hepa-1c1c7 cells. Oxidative DNA damage was estimated by nuclear condensation assessment, fluorescence-activated cell sorting analysis, and Comet assay. Rutaecarpine inhibited cell death induced by $500{\mu}M$ $H_2O_2$, as assessed by 4',6-diamidino-2-phenylindole (DAPI) staining. Treatment with rutaecarpine reduced the number of DNA strand breaks induced by $H_2O_2$, as assessed by DAPI staining and Comet assay, and increased quinone reductase, phosphatidylinositol 3-kinase, and pAkt protein levels, as assessed by western blotting.

A missense mutation in the coding region of the toll-like receptor 4 gene affects milk traits in Barki sheep

  • Sallam, Ahmed M.
    • Animal Bioscience
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    • v.34 no.4
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    • pp.489-498
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    • 2021
  • Objective: Milk production is one of the most desirable traits in livestock. Recently, the toll-like receptor (TLR) has been identified as a candidate gene for milk traits in cows. So far, there is no information concerning the contribution of this gene in milk traits in sheep. This study was designed to investigate the TLR 4 gene polymorphisms in Barki ewes in Egypt and then correlate that with milk traits in order to identify potential single nucleotide polymorphisms (SNPs) for these traits in sheep. Methods: A part of the ovine TLR 4 gene was amplified in Barki ewes, to identify the SNPs. Consequently; Barki ewes were genotyped using polymerase chain reaction-single strand conformation polymorphism protocol. These genotypes were correlated with milk traits, which were the daily milk yield (DMY), protein percentage (PP), fat percentage (FP), lactose percentage, and total solid percentage (TSP). Results: Age and parity of the ewe had a significant effect (p<0.05 or p<0.01) on DMY, FP, and TSP. The direct sequencing identified a missense mutation located in the coding sequence of the gene (rs592076818; c.1710C>A) and was predicted to change the amino acid sequence of the resulted protein (p.Asn570Lys). The association analyses suggested a significant effect (p<0.05) of the TLR genotype on the FP and PP, while the DMY tended to be influenced as well (p = 0.07). Interestingly, the presence of the G allele tended to increase the DMY (+40.5 g/d) and significantly (p<0.05 or p<0.01) decreased the FP (-1.11%), PP (-1.21%), and TSP (-7.98%). Conclusion: The results of this study suggested the toll-like receptor 4 (TLR4) as a candidate gene to improve milk traits in sheep worldwide, which will enhance the ability to understand the genetic architecture of genes underlying SNPs that affect such traits.

Base Specificity for DNA Interstrand Cross-Linking Induced by Anticancer Agent Bizelesin

  • Lee, Chong-Soon;Myung, Pyung-Keun;Gibson, Neil W.
    • Archives of Pharmacal Research
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    • v.19 no.3
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    • pp.191-196
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    • 1996
  • Bizelesin is a promising novel anticancer agent which is known to alkylate N3 of adenine to induce DNA interstrand cross-links (ISC) with in $5^I-TAATTA\; and\; 5^I-TAAAAAA$. We have investigated the base specificity for DNA ISC induced by bizelesin using oligomers containing the cross-linkable sequence $5^I-TAATTA\; and\; 5^I-TAAAAAA$. in which "N" was either A, C, G, or T. An analysis of denaturing polyacrylamide gel showed that bizelesin is able to induce DNA ISC in the duplex oligomer containing sequences $5^I-TAATTA\; and\; 5^I-TAAAAAA$. The formation of interstrand crosslinking did not occur in the sequences $5^I-TAATTA\; and\; 5^I-TAAAAAA$. DNA strand cleavage assay to determine the cross-linking site within $5^I-TAATTA$sequence showed that bizelesin alkylates guanine. These results demonstrate that bizelesin is able to induce DNA ISC at guanine but not at cytosine or thymine. In addition, guanine adducts have been found to be susceptible to DNA strand cleavage by exposure to hot piperidine. The extent of DNA strand cleavage, however, was not 100% efficient in either neutral pH buffer or hot piperidine.

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Terminal Nucleotide Sequences in the Double-stranded RNA Genome Segments of Infectious Pancreatic Necrosis Virus DRT Strain

  • Chung, Hye-Kyung;Park, Hong-Chul;Ichiro Uyeda;Masamichi Isogai;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.361-363
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    • 1996
  • The terminal regions of the double-stranded RNA (dsRNA) genome segments of infectious pancreatic necrosis virus (IPNV) DRT strain were sequenced. The dsRNAs, which were $^{32}P$-labelled at their 3'-termini by incubation with [$^{32}P$]pCp and T4 RNA ligase, were separated by 5$%$ polyacrylamide gel electrophoresis, and the segments A and B of IPNV-DRT were sequenced by two-dimensional gel electrophoresis. The 5'-terminal sequences of the IPNV-DRT plus strand from two genome segments were found to have the same conserved nucleotide (5'-CGG(C/A)A-), but the 3'-terminal sequences -CCCCAGGCG-3' and -CGGACCCCG-3' were found in the plus strand from segments A and B, respectively. The inverted oligonucleotide sequences of 3'-terminal of between segments A and B were found and they differ from those of other IPNVs.

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