• Title/Summary/Keyword: Endonuclease

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Isolation of Novel Non-Toxic Bacillus thuringiensis from Soil Samples in Korea (한국 토양으로부터 새로운 무독성 Bacillus thuringiensis 균주의 분리)

  • 노종열;박현우;김호산;진병래;강석권
    • Korean journal of applied entomology
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    • v.34 no.4
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    • pp.373-377
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    • 1995
  • Four Bacillus thuringiensis isolates obtained from soil samples in Korea produce parasporal inclusions non-toxic to 10 insect species of three orders, Lepidopera, Diptera and Coleoptera. These four isolates are named NTB-1, NTB-2, NTB-3 and NTB-4, respectively. The morphology of parasporal inclusions of four isolates observed by phase contrast- and scanning electron microscope was all ovoid. Characterization of four non-toxic B. thuringiensis isolates was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and restriction endonuclease analysis. The results showed that parasporal inclusion proteins and total plasmid DNA profiles of four isolates are different from other known non-toxic B. thuringiensis strains', suggesting that four isolates are novel.

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Regulation of Apoptosis by Nitrosative Stress

  • Kim, Ki-Mo;Kim, Peter K.M.;Kwon, Young-Guen;Bai, Se-Kyung;Nam, Woo-Dong;Kim, Young-Myeong
    • BMB Reports
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    • v.35 no.1
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    • pp.127-133
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    • 2002
  • Nitrosative stress can prevent or induce apoptosis. It occurs via S-nitrosylation by the interaction of nitric oxide (NO) with the biological thiols of proteins. Cellular redox potential and non-heme iron content determine S-nitrosylation. Apoptotic cell death is inhibited by S-nitrosylation of the redox-sensitive thiol in the catalytic site of caspase family proteases, which play an essential role in the apoptotic signal cascade. Nitrosative stress can also promote apoptosis by the activation of mitochondrial apoptotic pathways, such as the release of cytochrome c, an apoptosis-inducing factor, and endonuclease G from mitochondria, as well as the suppression of NF-${\kappa}B$ activity. In this article we reviewed the mechanisms whereby S-nitrosylation and nitrosative stress regulate the apoptotic signal cascade.

A Treatment of Basal cell Carcinoma on the Face of a Xeroderma Pigmentosum Patient: A Case Report (색소피부건조증 환자에서 발생한 안면부 기저세포암의 치료 증례 보고)

  • Bae, Jaehyun;Jun, Dongkeun;Kim, Jeenam;Lee, Myungchul;Shin, Donghyeok;Lim, So Dug;Choi, Hyungon
    • Korean Journal of Head & Neck Oncology
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    • v.36 no.1
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    • pp.39-44
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    • 2020
  • Xeroderma pigmentosum is a rare autosomal recessive disease, related to defects in DNA repair mechanism. It presents skin lesions on sun-exposed areas, leading to various skin cancer. Skin lesions can be treated with cryotherapy, skin resurfacing, 5-FU, Imiquimod, topical T4 endonuclease V, radiotherapy and genetic therapy, but invasive skin cancer should be treated by a surgery. We report a 12-year-old female xeroderma pigmentosum patient with recurrent basal cell carcinoma successfully treated by skin grafting. In that there is no cure for this disease, prevention and patient education is most important.

A Subpopulation of RNA3 of Cucumber mosaic virus Quasispecies

  • Park, Seung-Kook;Park, Sun-Hee;Yoon, Ju-Yeon;Park, Jang-Kyung;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.19 no.4
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    • pp.210-216
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    • 2003
  • This study examined the existence of genetically diverse population of Cucumber mosaic virus (CMV), known as quasispecies, from lily, Nicotiana benthamiana and from purified virions. Based on the conserved sequences of CMV lily isolates in intergenic region (IR) on RNA3, the genetic variation of IR from three different sources was investigated by a specific restriction endonuclease hydrolysis of amplified reverse transcription-polymerase chain reaction (RT-PCR) products using virus-specific primers, and was compared with IR sequences. The IR nucleotide sequences of CMV lily isolates were highly conserved, however, quasispecies was detected from all three sources in low level, containing sub-populations of RNA3. These subpopulations of RNA3 were inoculated onto zucchini squash by in vitro transcripts from corresponding full-length cDNA clones together with Eny RNA1 and 2 transcripts. The systemic symptom of zucchini plants infected by these quasispecies was chlorotic spotting, which was milder than severe mosaic and stunt symptom caused by Eny-CMV. The severity of symptom was correlated with RNA accumulation of viruses. These results suggest that the genome of CMV lily isolates consists of quasispecies populations.

DNA Methylation in Brain and Liver Tissues of Mice Infected with Scrapie Agent (스크래피에 감염된 마우스의 뇌 및 간조직에서의 DNA Methylation)

  • Choi, E.K.;Uyeno, S.;Ono, T.;Carp, R.I.;Kim, Y.S.
    • The Journal of Korean Society of Virology
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    • v.28 no.2
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    • pp.183-192
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    • 1998
  • DNA methylation degree in the several murine brain and liver genes of different ages and after scrapie infection have been examined by using methylation-sensitive restriction endonuclease digestion. We found that the methylation of c-fos and c-myc in the brain and liver was increased during the late fetal to one month postnatal developmental periods. However, those of the SGP-2, $S100{\beta}$, APP950, PrP, and APLP1 genes were decreased at the same periods. The comparison of the DNA methylation patterns between scrapie infected brains and controls demonstrated there is no significant difference in methylation degree of scrapie-infected brains. These observations indicate that DNA methylation might be importantly related to the aging process. The scrapie-infected murine brain was not significantly developed more senescent than the same age-controls did.

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Preliminary Research on the Expression, Purification and Function of the Apoptotic Fusion Protein, Sival

  • Zhang, Ya-Han;Yu, Lu-Gang;Zhu, Wan-Zhan;Wang, Sheng-Li;Wang, Dian-Dong;Yang, Yan-Xin;Yu, Xia
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8685-8688
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    • 2014
  • The objective of the present study was to investigate cloning, expression, and functions of the recombinant protein, Siva1. Siva1 gene was synthesized by RT-PCR from HCT116 cells. Plasmids were cleaved with the restriction endonuclease, BamH1/Sal1 and products were connected to pQE30, which underwent cleavage by BamH1/Sal1. The recombinant plasmid, pQE30-Siva1, was identified after digestion with restriction endonucleases followed by transformation into E. coli M15. Expression of Siva1 was induced by IPTG and identified by SDS-PAGE following purification with affinity chromatography. The results showed that size of Siva1 was 12 kDa, consistent with the molecular weight of the His-Siva1 fusion protein. Functional test demonstrated that Siva1 significantly inhibited the invasion and migration of HCT116 cells. It may thus find clinical application for control of cancers.

Direct Extraction of DNA from Soil for Amplification of 16S rRNA Gene Sequences by Polymerase Chain Reaction

  • Cho, Jae-Chang;Lee, Dong-Hun;Cheol, Cho-Young;Cho, Jang-Cheon;Kim, Sang-Jong
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.229-235
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    • 1996
  • Microgram quantities of DNA per gram soil were recovered with SDS- based and freeze-and thaw procedures. The average DNA fragment size was > 23 Kb. This method generated minimal shearing of extracted DNA. However, the DNA extracts still contained considerable amounts of humic impurities sufficient to inhibit PCR. Several approaches were used to reduce the interferences with the PCR (use of CTAF in extraction step, Elutip-d column purification, addition of BSA to PCR buffer) to accomplish PCR with DNA extract as a template. Most of the DNA extracts were not digested completely by restriction endonuclease, and CTAB-TREATED ane Elutip-d column purified DNA extracts were partially digested. Regarding as restriction enzyme digestion, all PCRs failed to amplify 16S rRNA gene fragments in the DNA extracts. In the case of DNA extracts only where BSA was added to PCR buffer, PCR was successfully conducted whether the DNA extracts were treated with CTAB or purified with columns. However, these two treatments were indispensable for humic impurity-rich DNA extracts to generate the PCR-compatible DNA samples. Direct extraction of DNA, coupled with these procedures to remove and relieve interferences by humic impurities and followed by the PCR, can be rapid and simple method for molecular microbiological study on soil microorganisms.

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Rapid Preparation of Truncated Transaminases using a PCR-based Cell-free Protein Synthesis System (PCR 기반의 무세포 단백질 발현 시스템을 이용한 절단 트랜스아미나제의 고속생산)

  • Kwon, Yong-Chan;Park, Kyung-Moon;Kim, Dong-Myung
    • KSBB Journal
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    • v.21 no.4
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    • pp.302-305
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    • 2006
  • In this work, we attempted the application of cell-free protein synthesis technology for the rapid generation of truncated enzymes. Truncated DNAs of a transaminase were PCR-amplified and directly expressed in cell-free protein synthesis reactions. Variants of the transaminase were rapidly prepared and analyzed for their enzymatic activity. Described method that combines the PCR and cell-free protein synthesis technologies will offer a versatile platform for the rapid generation of optimally modified protein species.

Molecular Cloning of Serratia marcescens Chitinase Gene into Escherichia coli (Serratia marcescens Chitinase 유전자의 대장균에로의 클로닝)

  • 장규일;김기석;조무제;이상열;신용철
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.129-135
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    • 1992
  • A chitinase gene of Serratia marcescens ATCC 27117 was cloned and expressed in Escherichiu di. A genomic library of S, marcescens was constructed with pUC 19 and screened using the swollen chitin agar plate for chitinolytic clones. A positive clone showing chitinclearance contains a recombinant pCHI 89, composed of 8.9 Kb chromosomal DNA fragment and pUC 19. Plasmid pCHI 89 produced 58 KD chitinase in E. coli, which was coincided with one of five extracellular chitinases produced by S. nzarccscens. Restriction endonuclease cleavage sites of the 8.9 Kb insert DNA fragment were mapped. E. coli JM109 harboring pCHI 89 inhibits the growth of a plant pathogenic fungus, Fusarium oxysporum.

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Identification of the Gene Products Responsible for F Plasmid Partitioning

  • Kim, Sung-Uk;Kazuo Nagai;Gakuzo Tamura;Yu, Ju-Hyun;Bok, Song-Hae
    • Journal of Microbiology and Biotechnology
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    • v.3 no.4
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    • pp.256-260
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    • 1993
  • DNA subfragments, sopA, sopB and sopC which help to maintain the stability of an ori C plasmid, were derived from a mini-F plasmid DNA (EcoRI restriction fragment f5) after digestion with restriction endonuclease, and cloned in the vector plasmid pBR322. The recombinant plasmids obtained were introduced into E. coli KY7231 and E. coli CSR603 strains, and proteins specified by the mini-F fragments were analysed by SDS-PAGE. Two proteins encoded by the F fragments were detected, and their molecular weights were 41,000 and 37,000 daltons. Fluorography after one and two dimensional gel electrophoresis of the lysates showed that these two proteins had been overproduced in the cells which were allowed to incorporate radioactive amino acid after plasmid amplification by chloramphenicol treatment. The isoelectric points of sopA and sopB proteins were 6.6 and 7.0, respectively.

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