• Title/Summary/Keyword: DNA density

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Identification of Two Entomopathogenic Bacteria from a Nematode Pathogenic to the Oriental Beetle, Blitopertha orientalis

  • Yi, Young-Keun;Park, Hae-Woong;Shrestha, Sony;Seo, Ji-Ae;Kim, Yong-Ook;Shin, Chul-Soo;Kim, Yong-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.968-978
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    • 2007
  • A pathogenic nematode, Butlerius sp., was isolated from Oriental beetle, Blitopertha orientalis. The infective juveniles exhibited dose-as well as time-dependent entomopathogenicity on the larvae of B. orientalis. Two bacterial species, Providencia vermicola (KACC 91278) and Flavobacterium sp. (KACC 91279), were isolated from the infective juveniles and identified. P. vermicola outnumbered Flavobacterium sp. in the nematode host, in which the colony density of P. vermicola was found to be 21 times higher than that of Flavobacterium sp. However, when the two bacterial species were cocultured in culture media without the nematode host, they showed similar growth rates. Both bacteria induced significant entomopathogenicity against Spodoptera exigua larvae infesting economically important vegetable crops, where P. vermicola was more potent than Flavobacterium sp.

An Efficient PEG/CaCl2-Mediated Transformation Approach for the Medicinal Fungus Wolfiporia cocos

  • Sun, Qiao;Wei, Wei;Zhao, Juan;Song, Jia;Peng, Fang;Zhang, Shaopeng;Zheng, Yonglian;Chen, Ping;Zhu, Wenjun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1528-1531
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    • 2015
  • Sclerotia of Wolfiporia cocos are of medicinal and culinary value. The genes and molecular mechanisms involved in W. cocos sclerotial formation are poorly investigated because of the lack of a suitable and reproducible transformation system for W. cocos. In this study, a PEG/CaCl2-mediated genetic transformation system for W. cocos was developed. The promoter Pgpd from Ganoderma lucidum effectively drove expression of the hygromycin B phosphotransferase gene in W. cocos, and approximately 30 transformants were obtained per 10 μg DNA when the protoplast suspension density was 106 protoplasts/ml. However, no transformants were obtained under the regulation of the PtrpC promoter from Aspergillus nidulans.

Present and Future Prospect in Zoysia Grass Breeding (한국잔디류(Zoysia Grass)의 육종현황)

  • 주영규;김두환;이재필;모숙연
    • Asian Journal of Turfgrass Science
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    • v.11 no.1
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    • pp.73-85
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    • 1997
  • Although zoysia grass (Zoysia spp.) has a lot of excellent chracteristics as warm-season grass, it have been limited in use due to slow establishment, low seed production, poor shade tolerance and other factors. Breeding trials have been continued from 1900's, much attentions have been paid especially in U.S.A., Korea and Japan recently. In U.S.A., more than 24 varieties had been evaluated at National Turfgrass Evaluation Program(NTEP) from 1991 to 1995 and some were regsistered as commercial. After the 6th International Turfgrass Research Conference at Japan in 1989, Japan Turfgrass In-corporation (JTI) sponsored by private companies and government carried out breeding programs for pest, salt, and shade tolerant and herbicide resistant varieties. JTI also has been trying to im-prove vigor and breed evergreen zoysia Korean breeders collected germplasms since 1960's. After USDA breeders came to Korean penesula in 1982, Korean breeders joined with USDA zoysia breeding project for several years. Many interspecific hybrids and natural selected varieties were breeded that period both in U.S.A and Korea. Breeding objectives were to extend green color period, improve leaf quality and density, and better leaf color at dormant stage. Since 1990's, zoysia grass breeding trials are getting more diverse in many points such as random amplified polymorphic DNA (RAPD) assay for ecotype identification. The objectives of this study are to evaluate germplasms in Korea, and also review the present status and future prospect in zoysia grass breeding in the world.

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Effect of the Dietary EPA in Replacement Diets for the Culture of Brackish Flea, Diaphanosoma celebensis (기수산 물벼룩의 배양을 위한 대체 먹이원 내 EPA 영향)

  • Park, Jin-Chul;Kwon, O-Nam;Park, Heum-Gi
    • Journal of Fisheries and Marine Sciences Education
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    • v.24 no.2
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    • pp.203-209
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    • 2012
  • We investigated the effect on dietary EPA (eicosapentaenoic acid) source (EP) into formulated diets for growth of brackish flea, Diaphanosoma celebensis. The highest density and specific growth rate (SGR) of brackish fleas were observed on Tetraselmis suecica (TE) trial, but these of trial were not significantly differed with that of Chlorella (CH) + Crypthecodinium sp. (CR) + EP trial, contained EPA oil (P>0.05). Contrastively, CH trial showed the lowest SGR. And in the RNA/DNA ratio, 0.08 of TE trial was the highest ratio out of whole trials, but the trial not significantly differed with that of CH + CR + EP trial (P>0.05). But, the ratio of CH trial was the lowest ratio at 0.05 out of whole trials (P<0.05). Through out results, the EPA source for flea culture was showed a positive effect through their growth and SGR. Therefore, we suggested that a usage of the source with CH could replace T. suecica as good diet for culture of the brackish flea.

Studies on the Efficient Separation of Primordial Germ Cells and Introduction of Foreign DNA in the Chicken (닭에서 원시생식세포의 효율적 분리 및 외래 유전자 전이에 관한연구)

  • 정동기;한재용
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 1999.11a
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    • pp.11-33
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    • 1999
  • This study was conducted to determine the embryonic stages for the isolation of the highest number of PGCs and to improve PGCs enrichment method. The primordial germ cells(PGCs) from different sources of chick embryos were isolated. The embryonic stage having the highest number of PGCs from each sources was selected ; 1-day-old embryos for germinal crescent (stage 6-8), 2.5-day-old embryos for blood (stage 17-18) and 5.5-day-old embryos for gonad (stage 27-28). The number of PGCs from one embryonic germinal crescent, blood and gonad was about 87$\pm$1.8, 103$\pm$4.0, and 932$\pm$10.9, respectively. The viability of PGCs after Ficoll from each sources was similar, showing approximately 70%. the PGCs enrichment method was improved using Ficoll density gradient centrifugation. After this step the purity of PGCs from germinal crescent, blood, and gonad was 45$\pm$9.10%, 85$\pm$1.18%, and 86$\pm$0.19%, respectively. Also, PGCs were picked up by mouth pipette to improve the purity. This improved method for the separation of PGCs from different sources will serve as a useful too to preserve the foundation stocks of poultry and to produce germline chimeras.

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Effect of single nucleotide polymorphisms on intramuscular fat content in Hungarian Simmental cattle

  • Anton, Istvan;Huth, Balazs;Fuller, Imre;Rozsa, Laszlo;Hollo, Gabriella;Zsolnai, Attila
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.9
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    • pp.1415-1419
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    • 2018
  • Objective: To estimate effect of single nucleotide polymorphisms on the intramuscular fat content (IMF) of Hungarian Simmental bulls. Methods: Genotypes were determined on high-density Illumina Bovine DNA Chip. After slaughtering of animals, chemical percentage of intramuscular fat was determined from longissimus dorsi muscle. A multi-locus mixed-model was applied for statistical analyses. Results: Analyses revealed four loci (rs43284251, rs109210955, rs41630030, and rs41642251) to be highly associated ($-{\log}_{10}P$>12) with IMF located on chromosome 1, 6, 13, and 17, respectively. The frequency of their minor alleles was 0.426, 0.221, 0.162, and 0.106. Conclusion: The loci above can be useful in selection programs and gives the possibility to assist selection by molecular tools.

A new cell-direct quantitative PCR based method to monitor viable genetically modified Escherichia coli

  • Yang Qin;Bo Qu;Bumkyu Lee
    • Korean Journal of Agricultural Science
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    • v.49 no.4
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    • pp.795-807
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    • 2022
  • The development and commercialization of industrial genetically modified (GM) organisms is actively progressing worldwide, highlighting an increased need for improved safety management protocols. We sought to establish an environmental monitoring method, using real-time polymerase chain reaction (PCR) and propidium monoazide (PMA) treatment to develop a quantitative detection protocol for living GM microorganisms. We developed a duplex TaqMan quantitative PCR (qPCR) assay to simultaneously detect the selectable antibiotic gene, ampicillin (AmpR), and the single-copy Escherichia coli taxon-specific gene, D-1-deoxyxylulose 5-phosphate synthase (dxs), using a direct cell suspension culture. We identified viable engineered E. coli cells by performing qPCR on PMA-treated cells. The theoretical cell density (true copy numbers) calculated from mean quantification cycle (Cq) values of PMA-qPCR showed a bias of 7.71% from the colony-forming unit (CFU), which was within ±25% of the acceptance criteria of the European Network of GMO Laboratories (ENGL). PMA-qPCR to detect AmpR and dxs was highly sensitive and was able to detect target genes from a 10,000-fold (10-4) diluted cell suspension, with a limit of detection at 95% confidence (LOD95%) of 134 viable E. coli cells. Compared to DNA-based qPCR methods, the cell suspension direct PMA-qPCR analysis provides reliable results and is a quick and accurate method to monitor living GM E. coli cells that can potentially be released into the environment.

A Theoretical Study for Estimation of Oxygen Effect in Radiation Therapy (방사선 조사시 산소가 세포에 미치는 영향의 이론적 분석)

  • Rena J. Lee;HyunSuk Suh
    • Progress in Medical Physics
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    • v.11 no.2
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    • pp.157-165
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    • 2000
  • Purpose: For estimation of yields of l)NA damages induced by radiation and enhanced by oxygen, a mathematical model was used and tested. Materials and Methods: Reactions of the products of water radiolysis were modeled as an ordinary time dependant equations. These reactions include formation of radicals, DNA damage, damage repair, restitution, and damage fixation by oxygen and H-radical. Several rate constants were obtained from literature while others were calculated by fitting an experimental data. Sensitivity studies were performed changing the chemical rate constant at a constant oxygen number density and varying the oxygen concentration. The effects of oxygen concentration as well as the damage fixation mechanism by oxygen were investigated. Oxygen enhancement ratio(OER) was calculated to compare the simulated data with experimental data. Results: Sensitivity studies with oxygen showed that DNA survival was a function of both oxygen concentration and the magnitude of chemical rate constants. There were no change in survival fraction as a function of dose while the oxygen concentration change from 0 to 1.0 x 10$^{7}$ . When the oxygen concentration change from 1.0 $\times$ 107 to 1.0 $\times$ 101o, there was significant decrease in cell survival. The OER values obtained from the simulation study were 2.32 at 10% cell survival level and 1.9 at 45% cell survival level. Conclusion: Sensitivity studies with oxygen demonstrated that the experimental data were reproduced with the effects being enhanced for the cases where the oxygen rate constants are largest and the oxygen concentration is increased. OER values obtained from the simulation study showed good agreement for a low level of cell survival. This indicated that the use of the semi-empirical model could predict the effect of oxygen in cell killing.

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Quantitative Real-time PCR using Lactobacilli as Livestock Probiotics (Real-time PCR을 이용한 가축생균제용 유산균 정량분석)

  • Choi, Yeon-Jae;Kim, Sun-Ho;Gu, Min-Jeong;Choe, Han-Na;Kim, Dong-Un;Cho, Sang-Bum;Kim, Su-Ki;Jeon, Che-Ok;Bae, Gui-Seok;Lee, Sang-Seok
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1896-1901
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    • 2010
  • This study was conducted using quantitative real-time PCR using Lactobacilli as probiotics. Quantitative real-time PCR (RT PCR) was conducted via a method involving SYBR Green 1 and a probe. Plasmid DNA was cloned using the 16S-23S rRNA intergenic species region. Gene clones were diluted from $10^2$ to $10^{10}$. Standard curves were constructed via Ct values obtained from the results of Real-time PCR via the aforementioned SYBR Green 1 and probe method. Plasmid DNA was also cloned using the 16S-23S rRNA intergenic species region and the gene clones were diluted from $10^2$ to $10^{10}$ copy numbers via the probe method. Using RT PCR, a standard curve of plasmid DNA copy numbers was also determined. The slope value for the Y-axis intercept and $R^2$ value were measured as -3.346, 33.18, and 0.993, respectively, via the first method. For the second method, the slope value for the Y-axis intercept and $R^2$ were -3.321, 31.10 and 0.995, respectively. The PCR inhibitor could not express the detection curve at a copy number over $10^{10}$ via either method, owing to high DNA density. The DNA extract from probiotics was diluted without pre-culturing, and 16 products were amplified via both methods. The Ct value was 11.06~18.12 in the first method and 16.74~22.11 in the second method. Measured probiotics and log copy values were largely similar among the methods used. It was concluded that both methods are effective for analysis, but further research will be required to verify the optimal method.

Study of Gene-gene Interaction within GNB3, ACE, ADRB3, ADRB2 among Korean Female Subject (한국인 비만 여성의 GNB3, ACE, ADRB3, ADRB2 유전자 다형성간의 상호관계에 관한 연구)

  • Choi Hyun;Bae Hyun su;Hong Moo chang;Shin Hyun Dae;Shin Min Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1426-1436
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    • 2004
  • There have been several reports on the relationship between G protein β3 subunit gene (GNB3), angiotensin converting enzyme gene (ACE), β3-adrenergic receptor gene (ADRB3), and β2-adrenergic receptor gene (ADRB2) genotype and obesity or obesity related disease. The objective of this study was to examine the relationship between the combinations of these four genes' polymorphism and probability of obesity related disease in Korean female subjects. The experimental group was consisted of 85 obese Korean female subjects (body mass index, BMI≥27㎏/㎡). To determine the polymorphism, genomic DNA was isolated, and PCR was performed. Serological examinations (fasting plasma glucose, FPG; aspartate aminotranferase, AST; alanine aminotransferase, ALT; total cholesterol, TC; triglyceride, TG; high density lipoprotein-cholesterol, HDL; low density lipoprotein-choles terol, LDL) were carried by an autoanalyzer and serological methods. BMI, waist circumference (WC), hip circumference and waist hip ratio (WHR) were measured. Consequencely in the analysis with grouping of general genotyping and variant allele carrier/non-carrier, the result was not significantly different within all gene combinations and polymorphic pairings except higher waist circumference in Arg16Arg group of ADRB2 codon16 (P=0.024). And there was no significantly contrast result about age, height, weight, AST and ALT that are index feature of liver and gall bladder disease in polymorphic pairings of gene combinations. However, the statistical analysis of waist-hip ratio and waist circumference that could be recognized as the physical type of obesity showed T-Arg16 pairing carrier in GNB3-ADRB2 codon16 combination had increased WHR and WC significantly (P=0.046 and P=0.015 respectively). Futhermore, the levels of total cholesterol (TC) and low density lipoprotein choresteral (LDL) were significantly lower in C-I pairing of GNB3-ACE combination (P=0.032 and P=0.005). These results suggest that the T-Arg16 pairing carrier in GNB3-ADRB2 codon16 gene might have increased waist circumference and C-I pairing carrier in GNB3-ACE combination have lower possibility of contraction of cardiovascular disease related cholesterol and LDL despite of obese state.