• Title/Summary/Keyword: Phage

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Selective Gene Transfer to Hepatocellular Carcinoma Using Homing Peptide-Grafted Cationic Liposomes

  • Tu, Ying;Kim, Ji-Seon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.821-827
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    • 2010
  • Gene delivery that provides targeted delivery of therapeutic genes to the cells of a lesion enhances therapeutic efficacy and reduces toxic side effects. This process is especially important in cancer therapy when it is advantageous to avoid unwanted damage to healthy normal cells. Incorporating cancer-specific ligands that recognize receptors overexpressed on cancer cells can increase selective binding and uptake and, as a result, increase targeted transgene expression. In this study, we investigated whether a peptide capable of homing to hepatocellular carcinoma (HCC) could facilitate targeted gene delivery by cationic liposomes. This homing peptide (HBP) exhibited selective binding to a human hepatocarcinoma cell line, HepG2, at a concentration ranging from 5 to 5,000 nM. When conjugated to a cationic liposome, HBP substantially increased cellular internalization of plasmid DNA to increase the transgene expression in HepG2 cells. In addition, there was no significant enhancement in gene transfer detected for other human cell lines tested, including THLE-3, AD293, and MCF-7 cells. Therefore, we demonstrate that HBP provides targeted gene delivery to HCC by cationic liposomes.

The Characterization of Mitochondrial DNA of Korean Ginseng (Panax ginseng C.A. Meyer) (고려인삼의 미토콘드리아 DNA의 분자생물학적 특성연구)

  • Lim, Yong-Pyo;Park, Kwang-Tae
    • Journal of Ginseng Research
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    • v.14 no.2
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    • pp.310-316
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    • 1990
  • This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular 9enetical approach of energy Production related mechanism in Panax ginseng. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. mtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not. Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRII treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN I and EcoRII. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector. Genomic library was screened and purified for further research including restriction mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned from the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

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Purification and Charaterization of Antimicrobial Peptide from Roots of Pokeweed (미국자리공(Phytolacca americana L.) 뿌리의 항균 펩타이드 정제 및 특성연구)

  • Kim, Jeong-Joo;Jang, Hye-Young;Kim, Jae-Ho
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.385-390
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    • 2003
  • An antimicrobial peptide was purified from the roots of Phytolacca americana L. and was designated as PAMP-r. Purification was carried out by DEAE-cellulose anion exchange, sephadex G-75 gel filtration, Mono S cation exchange, and Resource RPC reverse phage chromatography. The molecular weight of PAMP-r was estimated to be about 4,900 Da by 15% SDS-PAGE under reducing condition. PAMP-r exhibited a broad spectrum of antimicrobial activity. PAMP-r was stable against heat and pH treatment; its activity was not diminished by the heat treatment up to $80^{\circ}C$ for 30 min, and it showed a pH stability in the range between pH 3.0 to pH 8.0.

Recent Insights into Aeromonas salmonicida and Its Bacteriophages in Aquaculture: A Comprehensive Review

  • Park, Seon Young;Han, Jee Eun;Kwon, Hyemin;Park, Se Chang;Kim, Ji Hyung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1443-1457
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    • 2020
  • The emergence and spread of antimicrobial resistance in pathogenic bacteria of fish and shellfish have caused serious concerns in the aquaculture industry, owing to the potential health risks to humans and animals. Among these bacteria, Aeromonas salmonicida, which is one of the most important primary pathogens in salmonids, is responsible for significant economic losses in the global aquaculture industry, especially in salmonid farming because of its severe infectivity and acquisition of antimicrobial resistance. Therefore, interest in the use of alternative approaches to prevent and control A. salmonicida infections has increased in recent years, and several applications of bacteriophages (phages) have provided promising results. For several decades, A. salmonicida and phages infecting this fish pathogen have been thoroughly investigated in various research areas including aquaculture. The general overview of phage usage to control bacterial diseases in aquaculture, including the general advantages of this strategy, has been clearly described in previous reviews. Therefore, this review specifically focuses on providing insights into the phages infecting A. salmonicida, from basic research to biotechnological application in aquaculture, as well as recent advances in the study of A. salmonicida.

Analysis of DNA Conformation in the Particles of Bacteriophage P4 Mutant, P4 ash8 (박테리오파아지 P4 ash8 sid71 입자 내 DNA 형태 분석)

  • Song, Jae-Ho;Kim, Kyoung-Jin
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.62-66
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    • 2006
  • To study the packaging mechanism of the bacteriophage P2-P4 system which is a useful experimental tool for the study of viral capsid assembly, we analyzed the DNA contents of P4 sid- mutant, P4 ash8 sid71's phage particles. Two kind of particles having different density were separated by the CsCl buoyant equilibrium density gradient experiment with fresh made stock of P4 ash8 sid71. The DNA from each particles was prepared and its conformations was analyzed by electrophoresis. Unexpectedly, both particles contain not only dimeric and trimeric but also monomeric P4 DNA.

Isolation and Identification of Cyanophage from Eutrophic Water (부영양화 수역에서의 Cyanophage 의 분리와 동정)

  • Kim, Min;Choi, Yong-Keel
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.524-527
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    • 1992
  • Synechococcus sp. cyanophage was isolated from Baekwoon reservoir located in KyonggiDo. The cyanophage was purified by employing ultrafiltration. differential centrifugation. and sucrose density gradient centrifugation. Electron microscopic observation indicated that the sizes of its isometric head and contractile tail are 89 nm and] II nm. respectively. which means that the isolated cyanophage is included in the group. Myoviridae. The cyanophage maintained the stability of more than 50 percent from $20^{\circ}C$ to $40^{\circ}C$ and from pH 5 to 8. and had the maximal infectivity at $30^{\circ}C$ and pH 9 implying its ecological significance.

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Strain Identification of Xanthomonas oryzae by Bacteriophages and their Distribution in Korea (Bacteriophage에 의한 벼흰빛잎마름병균의 계통분류 및 그 분포에 관한 연구)

  • Cho Yong Sup;Park Chang Seuk;Choi Yong Chul;Lee Kyung Hee
    • Korean journal of applied entomology
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    • v.11 no.2
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    • pp.85-89
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    • 1972
  • Fifty-one isolates of Xanthomonas oryzae collected from all over the Korean paddy field were classified as the eight different strains by using four different bacteriophage types. Strain-F was the most prevalent form and strain-H,C, and A followed the strain-F, respectively. Strain-D, however, had not found even single isolate although the strain·D is the most prevalent form at Japan and southern countries of Asia. There was no corelation between the type of strains and the area front which the strain had been collected. Six different types of strains were isolated from IR 667 varieties whereas the three strains were maximum that could isolated from single variety other than IR 667.

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Construction and Characterization of Escherichia coli-Corynebacterium nephridii Hybrid Thioredoxins

  • Sa, Jae-Hoon;Kim, Kyung-Hoon;Lim, Chang-Jin
    • BMB Reports
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    • v.28 no.1
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    • pp.51-56
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    • 1995
  • Thioredoxin is a small redox protein with an active-site disulfide/dithiol, and is ubiquitous in bacteria, plants, and animals. To investigate the structure-function relationship of thioredoxin, the genes encoding Escherichia coli thioredoxin and Corynebacterium nephridii thioredoxin C3 were fused via a common restriction site in the nucleotide sequence coding for the active site of the proteins to generate two chimeric thioredoxins, designated E-C3(N to C-terminal) and C3-E. The hybrid thioredoxin genes were put under the T7 promoter and their productions were confirmed. The two hybrid thioredoxins complemented phenotypes of a thioredoxin-deficient E. coli strain. A strain containing the C3-E hybrid thioredoxin supported growth of the T7 phage, whereas a strain expressing the E-C3 hybrid thioredoxin did not. However, both hybrids supported growth of M13 phages. The two hybrid thioredoxins were also characterized in other aspects. Differences in activity between the hybrid thioredoxins were attributed to altered interactions of the N- and C-terminal domains of the molecule, which produced changes in the three-dimensional structure of the active site region.

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A Study on the Ecological Life Style in the Home and Preservation of the Earth Environment (지구환경보전과 가정에서의 생태적인 생활양식)

  • 최남숙
    • Hwankyungkyoyuk
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    • v.7 no.1
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    • pp.88-105
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    • 1994
  • Since the Rio Convention of 1992, the international community has displayed a heightened consciousness concerning the Earth environment and domestic attention on the said subject has also rapidly increased. An Ecological life style signifies the conscious efforts to preserve the environment interpreted into quotidien action. For the realization of conservation in action, an understanding of the specific details of environmental preservation must precede. The environmental conservation action in the homes was systematically approached in this study as an input-process-output and feedback cycle. The input phage is the consumer oriented buying stage, the while the process is the actual use of the products and the output stage consists of categorizing and producing wales, the feodbach being recycling of resources. At each stage the attitude which is devoted to environmental consciousness plays a big ride in the actual preservation of the Earth environment. in order to effectively conserve the environment, efforts displayed by the business sector is as important as implementation of environmentally conscious action at home. During the process of producing and distributing products, and attitude and action in preservation efforts are crncial and the responsibility ageat. For the business sector to effectively initiate environmental preservation into action, institutional support from the government and cooperation from the homes is of essence. Consolidated efforts to atter the superfrony life style into ecological life styles and ralues is essential to solve the most important socio-economic issue of the 90's of preserving the earth.

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MOLECULAR CLONING OF CHICKEN INTERFERON-GAMMA (닭 인터페론 유전자의 클로닝에 관한 연구)

  • ;Hyun Lillehoj
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 1999.11a
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    • pp.34-50
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    • 1999
  • A cDNA encoding chicken interferon-gamma (chIFN-${\gamma}$) was amplified from P34, a CD4$^{+}$ T-cell hybridoma by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pUC18. THe sequences of cloned PCR products were determined to confirm the correct cloning. Using this cDNA as probe, chicken genomic library from White Leghorn spleen was screened. Phage clones harboring chicken interferon-gamma (chIFN-${\gamma}$) were isolated and their genomic structure elucidated. The chIFN-${\gamma}$ contains 4 exons and 3 introns spanning over 14 kb, and follows the GT/AG rule for correct splicing at the exon/intron boundaries. The four exons encode 41, 26, 57 and 40 amino acids, respectively, suggesting that the overall structure of IFN-${\gamma}$ is evolutionairly conserved in mammalian and avian species. The 5’-untranslated region and signal sequences are located in exon 1. Several AT-rich sequences located in the fourth exon may indicate a role in mRNA turnover. The 5’-flanking region contains sequences homologous to the potential binding sites for the mammalian transcription factors, activator protein-1(AP-1) activator protein-2(AP-2) cAMP-response element binding protein(CREB), activating transcription factor(ATF), GATA-binding fator(GATA), upstream stimulating factor(USF), This suggests that the mechanisms underlying transcriptional regulation of chicken and mammalian IFN-${\gamma}$ genes may be similar.r.

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