• Title/Summary/Keyword: DnaJ

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New Antisense RNA Systems Targeted Against Plant Pathogens

  • Matousek, J.;Vrba, L.;Kuchar, M.;Pavingerova, D.;Orctova, L.;Ptacek, J.;Schubert, J.;Steger, G.;Beier, H.;Riesner, D.
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.379-385
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    • 2000
  • tRNA and 7SL RNA based antisense vehicles were prepared by inserting conserved anti-viral and anti-viroid domains. Anti-PVS coat protein leader sequence (ACPL) and antistructural antihairpin domain of PSTVd (AHII) were inserted in tRNA cassette; anti- zing finger domain of PVS, AHII and anti hop latent viroid ribozyme were inserted in 7SL RNA gene isolated from A. thaliana. These constructs were shown to be transcribed both, in in vitro and in in vivo conditions. However, it followed from our work that closely linked position of PoIII reference genes and PoIIII antisense genes within T-DNA lead to the impairment of RNA expression in transgenic plants. To assay in vivo transcription of antisense genes, hairy root potato cultures were established using h. tumefaciens A4-24 bearing both, Ri plasmid and PoIII-promoterless plant expression vectors with antisense RNA genes. Expression of antisense RNA in transgenic potato tissues was proven by specific RT-PCR reactions.

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An embryoprotective role for glucose-6-phosphate dehydrogenase in developmental oxidative stress and chemical teratogenesis

  • Nicol, Christopher J.;Zielenski, Julian;Tsui, Lap-Chee;Wells, Peter G.
    • Proceedings of the Korea Environmental Mutagen Society Conference
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    • 2002.05a
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    • pp.48-64
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    • 2002
  • The primary recognized health risk from common deficiencies in glucose-6-phosphate dehydrogenase (G6PD), a cytoprotective enzyme for oxidative stress, is red blood cell hemolysis. Here we show that litters from untreated pregnant mutant mice with a hereditary G6PD deficiency had increased prenatal (fetal resorptions) and postnatal death. When treated with the anticonvulsant drug phenytoin, a human teratogen that is commonly used in pregnant women and causes embryonic oxidative stress, G6PD-deficient dams had higher embryonic DNA oxidation and more fetal death and birth defects. The reported G6PD gene mutation was confirmed and used to genotype fetal resorptions, which were primarily G6PD deficient. This is the first evidence that G6PD is a developmentally critical cytoprotective enzyme for both endogenous and xenobiotic-initiated embryopathic oxidative stress and DNA damage. G6PD deficiencies accordingly may have a broader biological relevance as important determinants of infertility, in utero and postnatal death, and teratogenesis.-Nicol, C. J., Zielenski, J., Tsui, L.-C., Wells, P. G. An embryoprotective role for glucose-6-phosphate dehydrogenase in developmental oxidative stress and chemical teratogenesis.

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Identification of the Differentially Expressed Genes of Hanwoo During the Growth Stage by Subtractive cDNA Hybridization (Subtraction 기법을 이용한 한우 성장 단계 특이 발현 유전자 탐색)

  • Jang, Y.S.;Kim, T.H.;Yoon, D.H.;Park, E.W.;Cheong, I.C.;Jo, J.K.
    • Journal of Animal Science and Technology
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    • v.44 no.1
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    • pp.13-22
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    • 2002
  • To identify the differentially expressed genes at growth stage of Hanwoo, we constructed the subtractive cDNA library from loin mRNA of 12- and 24-month old Hanwoo by PCR-based subtraction. The fourteen genes were confirmed by sequencing and reverse northern blot analysis, and they were selected as candidate of putative genes differentially expressed at the growth stage of Hanwoo. Three subtracted cDNA fragments that expressed specific signal to cDNA probe for 6-month-old loin of Hanwoo were highly homologous to those of the genes encoding EPV 20, Ca2+ATPase, and TCTP, respectively. The nine cDNA clones showed intense signal to cDNA probe from 12-month-old loin of Hanwoo, and highly homologus to those of genes encoding VCP, HSP 70, aldolase A, MSSK1, GM-2 activator protein, ryanodine receptor, acidic ribosomal phosphoprotein p1, ADP/ATP translocase, and UCP 2, respectively. Two subtracted cDNA clones that expressed specific signal to cDNA probes for 12- and 24-month-old loin of Hanwoo were detected. One of them was highly homologus to the gene encoding ferrochelatase and the other was highly homologus to the gene encoding ADRP.

Genetic Diversity and Phylogenetic Analysis of South-East Asian Duck Populations Based on the mtDNA D-loop Sequences

  • Sultana, H.;Seo, D.W.;Bhuiyan, M.S.A.;Choi, N.R.;Hoque, M.R.;Heo, K.N.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.12
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    • pp.1688-1695
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    • 2016
  • The maternally inherited mitochondrial DNA (mtDNA) D-loop region is widely used for exploring genetic relationships and for investigating the origin of various animal species. Currently, domestic ducks play an important role in animal protein supply. In this study, partial mtDNA D-loop sequences were obtained from 145 samples belonging to six South-East Asian duck populations and commercial duck population. All these populations were closely related to the mallard duck (Anas platyrhynchos), as indicated by their mean overall genetic distance. Sixteen nucleotide substitutions were identified in sequence analyses allowing the distinction of 28 haplotypes. Around 42.76% of the duck sequences were classified as Hap_02, which completely matched with Anas platyrhynchos duck species. The neighbor-joining phylogenetic tree also revealed that South-East Asian duck populations were closely related to Anas platyrhynchos. Network profiles were also traced using the 28 haplotypes. Overall, results showed that those duck populations D-loop haplotypes were shared between several duck breeds from Korea and Bangladesh sub continental regions. Therefore, these results confirmed that South-East Asian domestic duck populations have been domesticated from Anas platyrhynchos duck as the maternal origins.

The Correlation between DNA Ploidy and the Clinicohistologic Findings in Colorectal Cancer (대장직장암에서 임상조직학적 소견과 DNA ploidy와의 상관관계)

  • Lee Suk Ho;Kim Hun Jung;Kim Woo Chul;Cho Young Kap;Loh John J K;Woo Ze Hong;Hwamg Tae Sook
    • Radiation Oncology Journal
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    • v.18 no.3
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    • pp.187-193
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    • 2000
  • Purpose : DNA ploidy pattern was shown to correlate with several clinicohistologic findings in several tumors. Aim of this study was to evaluate the correlation of the clinicohistologic findings in colorectal cancer and the failure pattern in rectosigmoid cancer with DNA ploidy. Materials and Methods DNA flow cytometry using the Hedley methods on paraffin embedded specimen from 117 patients with colorectal cancers after curative resection was peformed. We tried to find the correlation between DNA ploidy and various clinicohistologic findings. And then the correlation DNA ploidy and the failure pattern in 75 patients of rectosigmoid cancer was analized. Results : Forty samples (34.2%) from tumors gave aneuploidy histogram. There was no significant difference in the frequency of DNA aneuploidy in terms of age, sex, depth of invasion, location and Dukes stage. But there was a significant correlation between DNA ploidy and the failure rates in Dukes stage B rectosigmoid cancer (P=0.048). Conclusions : These findings suggest that DNA ploidy pattern shows the correlation with the treatment failure rates in Dukes stage B rectosigmoid, but not with many other clinicohistologic findings. However,more patients will be needed to disclose these findings.

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내열성 포도당 이성화효소를 생산하는 Streptomyces chibaensis J-59의 분리 및 동정

  • 주길재;권기석;이인구
    • Microbiology and Biotechnology Letters
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    • v.25 no.1
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    • pp.15-22
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    • 1997
  • A bacterial strain J-59 was isolated from a humus soil, which produced simultaneously a thermostable glucose isomerase as well as xylanase. The morphological, cultural and physiological characteristics of the isoisomerase strain J-59 were detemined by the use of the media and methods described in International Streptomyces Project. The chemotaxonomic characteristics of the isolated strain J-59 were determined by the analysis of G+C molar % of DNA, diaminipimelic acid, composition of fatty acid and menaquinone. As the results of various examinations, the strain J-59 was identified to be Streptomyces chibaensis. This strain produced glucose isomerase intracellularly and xylanase extracellularly when grown in a medium containing xylan, but it was not able to utilize the xylose or xylan as a carbon source. The glucose isomerase of S. chibaensis J59 was highly thermostable, which retained more than 75% activity in the presence of Co$^{2+}$ at 80$\circ $C for 72 h.

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Development and Evaluation of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Tylenchulus semipenetrans Using DNA Extracted from Soil

  • Song, Zhi-Qiang;Cheng, Ju-E;Cheng, Fei-Xue;Zhang, De-Yong;Liu, Yong
    • The Plant Pathology Journal
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    • v.33 no.2
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    • pp.184-192
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    • 2017
  • Tylenchulus semipenetrans is an important and widespread plant-parasitic nematode of citrus worldwide and can cause citrus slow decline disease leading to significant reduction in tree growth and yield. Rapid and accurate detection of T. semipenetrans in soil is important for the disease forecasting and management. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed to detect T. semipenetrans using DNA extracted from soil. A set of five primers was designed from the internal transcribed spacer region (ITS1) of rDNA, and was highly specific to T. semipenetrans. The LAMP reaction was performed at $63^{\circ}C$ for 60 min. The LAMP product was visualized directly in one reaction tube by adding SYBR Green I. The detection limit of the LAMP assay was $10^{-2}J2/0.5g$ of soil, which was 10 times more sensitive than conventional PCR ($10^{-1}J2/0.5g$ of soil). Examination of 24 field soil samples revealed that the LAMP assay was applicable to a range of soils infested naturally with T. semipenetrans, and the total assay time was less than 2.5 h. These results indicated that the developed LAMP assay is a simple, rapid, sensitive, specific and accurate technique for detection of T. semipenetrans in field soil, and contributes to the effective management of citrus slow decline disease.

APPLICATION OF RANDOMLY AMPLIFIED POLYMORPHIC DNA(RAPD) ANALYSIS METHOD FOR CLASSIFICATION AND BREEDING OF THE KOREAN GINSENG

  • Lim Y.P.;Shin C.S.;Lee S.J.;Youn Y.N.;Jo J.S.
    • Proceedings of the Ginseng society Conference
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    • 1993.09a
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    • pp.138-142
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    • 1993
  • Korean ginseng has been widely used as medicine from ancient times in Asia. Current breeding efforts in Korea include the individual plant selection and the subsequent pure - line isolation, and considerable number of lines with desirable traits have thus been isolated. However, there were rare data on genetic maker and its analysis for selection of superior varieties. For taxonomic characterization and development of genetic markers for ginseng breeding, molecular biological methods including the RFLP and RAPD methods were applied. Cytoplasmic DNA of ginseng was analyzed for RFLP analysis. However. there is no different pattern among the chloroplast DNA or mitochondrial DNA of variants. In the case of RAPD analysis, the band patterns using 4 of 10 RAPD primers show the distinctive polymorphism among 9 ginseng variants, and lines, and Similarity Index(SI) on polymorphism was calculated for the extent and nature of these variabilities in ginseng. The sequences of 4 selected primers were TGCCGAGCTG, AATCGGGCTG. GAAACGGGTG, and GTGACGTAGG. By SI based on the polymorphic band patterns, Chungkyung - Chong and Hwangskoog - Chong, and JakyungChong 81783 and Jinjakyung of Russia showed the most close SI. The data of KG10l coincided with the fact that it was released from Hwangskoog - Chong. and Jakyung - Chong 81783 and Jinjakyung of Russia showed the most close SI. The data of KG101 coincided with the fact that it was released from Hwangskoog - Chong by breeding process. The data of Jakyung strains indicated the significant variation among the strains. From these results, RAPD analysis method could be succesively applied to the classification and genetic analysis for breeding of Korean ginseng.

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Electrical transport characteristics of deoxyribonucleic acid conjugated graphene field-effect transistors

  • Hwang, J.S.;Kim, H.T.;Lee, J.H.;Whang, D.;Hwang, S.W.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.482-483
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    • 2011
  • Graphene is a good candidate for the future nano-electronic materials because it has excellent conductivity, mobility, transparency, flexibility and others. Until now, most graphene researches are focused on the nano electronic device applications, however, biological application of graphene has been relatively less reported. We have fabricated a deoxyribonucleic acid (DNA) conjugated graphene field-effect transistor (FET) and measured the electrical transport characteristics. We have used graphene sheets grown on Ni substrates by chemical vapour deposition. The Raman spectra of graphene sheets indicate high quality and only a few number of layers. The synthesized graphene is transferred on top of the substrate with pre-patterned electrodes by the floating-and-scooping method [1]. Then we applied adhesive tapes on the surface of the graphene to define graphene flakes of a few micron sizes near the electrodes. The current-voltage characteristic of the graphene layer before stripping shows linear zero gate bias conductance and no gate operation. After stripping, the zero gate bias conductance of the device is reduced and clear gate operation is observed. The change of FET characteristics before and after stripping is due to the formation of a micron size graphene flake. After combined with 30 base pairs single-stranded poly(dT) DNA molecules, the conductance and gate operation of the graphene flake FETs become slightly smaller than that of the pristine ones. It is considered that DNA is to be stably binding to the graphene layer due to the ${\pi}-{\pi}$ stacking interaction between nucleic bases and the surface of graphene. And this binding can modulate the electrical transport properties of graphene FETs. We also calculate the field-effect mobility of pristine and DNA conjugated graphene FET devices.

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Overproduction of the Escherichia coli Chaperones GroEL-GroES in Rhodococcus ruber Improves the Activity and Stability of Cell Catalysts Harboring a Nitrile Hydratase

  • Tian, Yuxuan;Yu, Chen, Huimin;Shen, Zhongyao
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.337-346
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    • 2016
  • Three combinations of molecular chaperones from Escherichia coli (i.e., DnaK-DnaJ-GrpE-GroEL-GroES, GroEL-GroES, and DnaK-DnaJ-GrpE) were overproduced in E. coli BL21, and their in vitro stabilizing effects on a nitrile hydratase (NHase) were assessed. The optimal gene combination, E. coli groEL-groES (ecgroEL-ES), was introduced into Rhodococcus ruber TH3. A novel engineered strain, R. ruber TH3G was constructed with the native NHase gene on its chromosome and the heterologous ecgroEL-ES genes in a shuttle plasmid. In R. ruber TH3G, NHase activity was enhanced 37.3% compared with the control, TH3. The in vivo stabilizing effect of ecGroEL-ES on the NHase was assessed using both acrylamide immersion and heat shock experiments. The inactivation behavior of the in vivo NHase after immersion in a solution of dynamically increased concentrations of acrylamide was particularly evident. When the acrylamide concentration was increased to 500 g/l (50%), the remaining NHase activity in TH3G was 38%, but in TH3, activity was reduced to 10%. Reactivation of the in vivo NHases after varying degrees of inactivation was further assessed. The activity of the reactivated NHase was more than 2-fold greater in TH3G than in TH3. The hydration synthesis of acrylamide catalyzed by the in vivo NHase was performed with continuous acrylonitrile feeding. The final concentration of acrylamide was 640 g/l when catalyzed by TH3G, compared with 490 g/l acrylamide by TH3. This study is the first to show that the chaperones ecGroEL-ES work well in Rhodococcus and simultaneously possess protein-folding assistance functions and the ability to stabilize and reactivate the native NHases.