• Title/Summary/Keyword: Agarose Gel

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Species identification of the Anopheles kyrcanus complex found in Korea using PCR (PCR을 이용한 우리나라에서 발견되는 얼룩날개모기속 모기의 종 동정)

  • Yong, Tae-Sun;Lee, Han-Il;Lee, In-Yong;Lee, Jong-Won;Hwang, Ui-Uk
    • Journal of Korea Association of Health Promotion
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    • v.4 no.1
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    • pp.68-74
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    • 2006
  • For identification of four sibling species of the Anopheles hyrcanus complex found in Korea, the 5.8 rDNA-ITS2-28S rDNA region of each species was sequenced and the species-specific primers wee designed The amplified PCR products obtained from each species were analyzed by agarose gel electrophoresis. The result showed a single species- specific band, I.e. 559bp, 432bp, 322bp and 192bp for An. sinensis, An. sp., An. lesteri and An. pullus, respectively. In conclusion, the species-specific PCR primers designed from ITS2 variable regions functioned successfully and specifically, and can be applied as a useful tool for identifying species of the Anopheles hyrcanus complex found in Korea.

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Circular Plasmid DNA from a Red Algae, Porphyra tenera (양식 참김(Porphyra tenera)에서 분리한 Circular Plasmid DNA)

  • 류태형;최학선;최경희;이춘환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.6
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    • pp.1160-1165
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    • 1998
  • When total cellular DNA was isolated from Porphyra tenera by ultracentrifugation on Hoechst dye/CsCl gradients method, plasmid like DNA's were concentrated at the upper band which were characterized with a A+T rich organelle DNA's in the CsCl gradients. Based on their electrophoretic migration in different concentration of agarose gel, buffer system, and electric power etc. and the results of restriction digestion, the plasmid like DNA's were concluded to have circular conformation. This is the first report of putative circular plasmid DNA from the P. tenera, which is a autonomously replicating plasmid existing with a high copy number plasmid in the cell. The minimum size of this plasmid estimated by restriction endonuclease digestion was appeared to be 2.5kb in size.

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Chrysanthemum stunt viroid in Dendranthema grandiflorum

  • Chung, Bong-Nam;Park, Gug-Seoun;Kim, Hyun-Ran;Kim, Jeong-Soo
    • The Plant Pathology Journal
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    • v.17 no.4
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    • pp.194-200
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    • 2001
  • Chrysanthemum stunt viroid (CSVd) ws identified in chrysanthemum cv. Chunkwang showing symptoms of stunt with leaf distortion (K1) and stunt with chlorosis of leaves (K2) collected from the main cultivation area of Masan, Kyongnam province in Korea. The specific RNAs related with the diseased chrysanthemums were detected. Full-length 354 bp CSVd cDNAs were amplified from infected tissue by reverse transcription and polymerase chain reaction using a pair of primers specific for CSVd sequence. The amplified cDNA products were analyzed by agarose gel electrophoresis and the specific cDNAs were cloned. Nucleotide sequences of the two CSVd isolates K1 and K2 varied. Phylogenetic analysis of the nucleotide sequences of CSVd isolates indicated that K1 was closely related with J2 and Am 2 isolates. K1 and K2 were transmitted by grafting to Dendranthema grandiflorum cv. Mistletoe, Gynura aurantiaca, and Lycopersicon esculentum cv. Rutgers. This is the first report of CSVd in D. grandiflorum in Korea.

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Detection of Salmonella typhi by Loop-mediated Isothermal Amplification Assay

  • Jo, Yoon-Kyung;Lee, Chang-Yeoul
    • Biomedical Science Letters
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    • v.14 no.2
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    • pp.115-118
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    • 2008
  • Salmonella typhi is frequent causes of foodborne illness and its detection is important for monitoring disease progression. In this study, by using general PCR and novel LAMP (Loop Mediated Isothermal Amplification) assay, we evaluated the usefulness of LAMP assay for detection of Salmonella typhi. In this LAMP assay, forward inner primer (FIP) and back inner primer (BIP) was specially designed for recognizing target invA gene. Target DNA was amplified and visualized as ladder-like pattern of bands on agarose gel within 60 min under isothermal conditions at $65^{\circ}C$. When the sensitivity and reproducibility of LAMP were compared to general PCR, there was no difference of reproducibility but sensitivity of LAMP assay was more efficient than PCR (the detection limit of LAMP assay was 30 fg, while the PCR assay was 3 pg). These results indicate that the LAMP assay is a potential and valuable means for detection of Salmonella typhi, especially for its rapidity, simplicity and low cost.

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Cloning and Expression of B. Aphaericus insecticidal toxin gene in E. coli

  • Lim, Pyong-Ok;Lee, Hong-Sup;Lee, Hyung-Hoan
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.515.2-515
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    • 1986
  • B. sphaericus 1593 K-5 synthesize a potent entomicidal toxin against mosquito larvae. B. sphaericus EcoRl DNA fragment carrying the biocidal activity was clonied and expression in E. coli JM83. For the construction of a recombinant plasmid bearing the toxin activity the DNA of B. sphaericus was partially digested by the EcoRl. The EcoRl DNA fragments were ligated to plasmid pUC 8-EcoR1 site. The transformants were selected on LB plates containing X-gall and ampicilline. The transformants were bioassayed against mosquito larvae of which two clones showed biocidal activity. The two clones were redigested with Eco R1 and analyzed by 0.7% agarose gel electrophorsis.

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Studies on the Endocrine Disruption in Wildlife Fish (어류생체지표를 이용한 내분비계장애 연구)

  • 구자민;류지성;정규혁;이철우;박응로;박광식
    • Environmental Analysis Health and Toxicology
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    • v.16 no.4
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    • pp.197-204
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    • 2001
  • Endocrine disruption in crucian carp (Carassius auratus) living in the branch of Han River were examined. Vitellogenin level in plasma was measured using ELISA system and aromatase mRNA level in brain was observed using RT-PCR technique. In all female fish, vitellogenin levels were in the range of 20∼40 $\mu\textrm{g}$/ml and aromatase mRNA expression could be detected on the agarose gel after RT-PCR. However, in case of males, vitellogenin level was elevated in only one fish, while vitellogenin was hardly detected in others. Aromatase was expressed in all males although the levels were relatively lower than the level in female fish. Testis-ova and any other histological changes of reproductive organ were not shown in both sexes.

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Characterization of Endogeneous Plasmids from Two Bacillus Isolates (Bacillus 속 분리균 2종의 내재형 Plasmids 특성분석)

  • 윤기홍
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.364-369
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    • 1999
  • In order to obtain the suitable plasmids for constructing plasmid vectors of Bacillus species, endogeneous plasmid DNAs were screended from thermo-tolerant soil bacteria. Based on agarose gel electrophoresis patterns of the isolated plasmid DNAs, two strains harboring small-size plasmids were selected. The isolated were identified to belong to the genus Bacillus on the basis of their morphological and biochemical properties, and named Bacillus sp. 3-3 and 77-8, respectively. The restriction endonuclease maps were determined for four plasmids including two plasmids from each Bacillus isolates. It is interesting that Bacillus sp. 3-3 and 77-8 have an identical plasmid according to the restriction maps. The three kinds of hybrid plasmids constructed by introducing each plasmid of two isolates into a Escherichia coli plasmid vector. pUCCm18 containing chloramplenicol resistance gene active in Bacillus strains, could be replicated in B. subtilis and B. licheniformis. These plasmids are very stable in B. subtilis, suggesting that the Bacillus plasmids identified in this work would be useful for development of new cloning vectors for Bacillus strains.

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Characteristics of Pseudomonas sp. degrading 2-methyl-4-chlorophenoxyacetic acid (2-메틸-4-클로로페녹시 아세트산을 분해하는 Pseudomonas 균주의 특성)

  • 은성호;박영두;이영록
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.389-393
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    • 1986
  • From the soil and river samples, some bacterial strains degrading chlorinated aromatic hydrocarbons were isolated and identified. Of the isolates, seven strains of Pseudomonas sp. harbouring plasmids were selected for their prominent degradative ability to 2-methyl-4-chlorophenoxyacetic acid. By agarose gel electrophoresis and curing experiment it was found that the genes for 2-methyl-4-chlorophenoxyacetic acid degradaiton were encoded on the plasmids in these selected strains. Antobiotic resistance and degradative ability for other herbicides of the strains were tested.

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Bacterial Virus DNA Damage Caused by Fumonisin B1 (Fumonisin B1에 의한 세균바이러스 DNA손상)

  • 이길수;조성국
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.34-38
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    • 1999
  • Fumonisin B1 is a secondary metabolite of Fusarium moniliforme, a contaminant of corn and corn product. Fumonisin B1 has been shown to be responsible for major toxicological effects of the fungus in rats, horses, and pigs. Fumonisin B1 induced λ DNA fragmentation, which was increased with incubation time, reducing agent NADPH and metal ion (Cu2+). The DNA damage was inhibited by dimethyl sulfoxide (DMSO) or mannitol as radical scavenger for free radicals. DNA fragmentation, induced by fumonisin B1 in the presence of 1 mM NADPH and 0.1 mM CuCl2, was inhibited by 100 mM DMSO. By the in vitro reaction of fumonisin B1 with supercoiled plasmid pBR322 DNA, plasmid DNA was relaxed, eventually linearized in the agarose gel electrphoresis. From rifampicin sensitive E. coli CSH138 in bacterial mutagenesis system, the rifampicin resistant E. coli mutants were obtained by fumonisin B1. These results suggest that fumonisin B1 may be a possible environmental mutagen in bacterial mutagen assay system.

Induction of FSTL1 Gene in Rat Myometrium by Exogenous Estrogen

  • Kim Yoon-Sik;Kim Yeon;Bae Hyung-Joon
    • Biomedical Science Letters
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    • v.10 no.4
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    • pp.479-483
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    • 2004
  • The aim of this study was carried to the induction of follistatin-like 1 (FSTL1) in rat myometrium tissue by exogenous estrogen. PCR was performed by using estrogen treatment after ovariectomy of myometrium mRNA and only ovariectomy of myometrium mRNA and on ovariectomy of myometrium mRNA in rat normal uterus tissue. The PCR products were visualized on agarose gel (1.2%) electrophoresis. FSTL1 mRNA expression level was significantly higher (**P<0.001 or *P<0.05) in estrogen treatment after ovariectomy than in only ovariectomy, and was significantly higher (**P<0.001) in no ovariectomy than in estrogen with treatment after ovariectomy. In conclusion, Although the mechanisms of FSTL1 gene were uncertain, It also might be inducted by exogenous estrogen. And also it is thought to be related to the estorgen mechanism.

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