• Title/Summary/Keyword: rDNA

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Polymorphisms of the Exons 13, 15 and 16 of Transferrin Gene in Cheju Horses (제주마 Transferrin Gene Exon 13, 15 및 16의 다형현상)

  • Kim, N.Y.;Lee, S.S.;Yang, Y.H.
    • Journal of Animal Science and Technology
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    • v.44 no.4
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    • pp.391-398
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    • 2002
  • This study was conducted to determine the polymorphism of transferrin exons 13, 15 and 16 by Single-Strand Conformation Polymorphism(SSCP) analysis and to compare their genotypes of Cheju horse Group I (Cheju Institute), Cheju horse Group II (farms), and Thoroughbred (KRA). SSCP of transferrin exon 13, 15, and 16 showed two (A, B), three (A, B, C) and three (A, B, C) codominant alleles, respectively. The Group I and Thoroughbred showed the similar frequencies of allele A and B in transferrin exon 13, but only allele A was observed in Group Ⅱ. In transferrin exons 15 and 16, the frequencies of each allele were different in each Groups. The multiple allele frequencies in exons 15 and 16 suggested that the genotyping of this locus could be used to identify an individual and to test the parentage of offspring. The probability for parentage exclusion were 0.46 and 0.374 for exons 15 and 16 for Cheju horse Group I. Among the 13 combined genotypes of exons 13, 15 and 16, the genotype AA-AB-AB (0.372) is the most common in Cheju horse Group I, but genotype AA-AA-AA is common in the Cheju horse Group II (0.366) and Thoroughbred (0.767). The present study showed two new SNP, which was at the cDNA position 1626 (A/G) in B allele of the exon 13 and 2075 (C/T) in C allele of the exon 16 resulting in amino acid change (Threonine $\longrightarrow$ Methionine). Result showed that polymorphism of exons 13, 15 and 16 in Cheju horses was as high as in Thoroughbred and there was a differences of transferrin allele frequencies in Cheju horses.

Effects of Cover Plants on Soil Microbial Community in Organic Apple Orchards (피복작물이 유기 사과과원 토양미생물상에 미치는 영향)

  • Oh, Young-Ju;Kang, Seok-Boem;Song, Yang-Ik;Choi, Jin-Ho;Paik, Woen-Ki
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.5
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    • pp.822-828
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    • 2012
  • Organic fruit production has increased due to consumer's interest and government's political support for environmentally-friendly agriculture. The aim of this study was to investigate the effects of cover plants on soil microbial community and establish the fruit cultivation method by organic farming techniques. Cover plants used as an organic nutrient source in an apple orchard were rye and barley, the Gramineae and red clover and hairy vetch, the Leguminosae. In the effects of cover plants on the soil chemical characteristics, the soil pH values were higher than that of conventional organic pear orchard. The content of P showed no significant difference between control and cover plant plots. Organic matter level was similar in control and Gramineae cover plant plots, while organic matter content in cover plants belong to Leguminosae was lower than that of control plot. K content was lower in the plots treated with rye and red clover than control plot, while K content in hairy vetch treated plot was higher than control plot. Ca content was lower in control plot than in cover plant treated plots. Concentrations of Mg in the plots treated with barley and hairy vetch was lower than control plot. In August rye and red clover covered soil showed higher bacterial community density than that of control soil and barley treated soil showed highest Actinomycetes community density among treatments. Barley and hairy vetch soils showed higher level of fungi community density than that of control soil in August. In pyrosequencing analysis barley treated soil showed highest distribution ratio of Actinomycetes among treatment. Our findings might be used as basic data for choosing cover plant with effective organic matter decomposition and nutrition supply capacity.

Construction and characterization of heterozygous diploid Escherichia coli (2배체 대장균의 제조와 그 특성)

  • Jung, Hyeim;Lim, Dongbin
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.406-414
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    • 2016
  • Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.

Microbial Community Analysis of Tarak, a Fermented Milk Product (우리나라 전통 발효유 타락의 미생물 균총 분석)

  • Lim, Goo-Sang;Lee, Kyung-Soo;Jang, Hye-Jin;Jung, Jin-Kyung;Lim, Ji-Young;Chun, TaeHoon;Han, Young-Sook;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1109-1114
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    • 2013
  • Microbial community analysis was performed on Tarak, a traditional Korean fermented milk product, by 16S rDNA cloning and pyrosequencing to obtain basic data for the standardization and systematization of the Tarak manufacturing process. Microbial analysis of the prokaryotic community revealed a slight difference in microbial abundance between Bontarak (n) and Tarak (n+1), but Firmicute was dominant at the phylum level. At the genus level, the Lactobacillus and Leuconostoc genera constituted over 90% of the population in Bontarak, but Lactococcus was the dominant genus in Tarak. Bontarak and Tarak showed further differences at the species level. Leuconostoc citreum was the dominant species in Bontarak, constituting 40% of the population. In eukaryotic community analysis, all samples were composed of Ascomycota at the phylum level. At the genus level, Saccharomyces was dominant in Bontarak (85% of the population), while Issatchenkia was dominant in Tarak (95% of the population). At the species level, Saccharomyces cerevisiae was detected at a relative abundance in Bontarak (82%), and Pichia kudriavzevii was the dominant species in Tarak, with a relative abundance of 95%. Sensory evaluation indicated that Tarak had a better appearance and texture than Bontarak. As sweetness was not significantly different between the two samples just slightly higher in Tarak, this was likely due to a significant decrease in sourness in Tarak. These results suggest that the microbial community used affects the quality of Tarak produced. Thus, a stable microbial community must be maintained for the production of Tarak with consistent quality.

A Dynamic Behavior Evaluation of the Curved Rail according to Lateral Spring Stiffness of Track System (궤도시스템의 횡탄성에 따른 곡선부 레일의 동적거동평가)

  • Kim, Bag-Jin;Choi, Jung-Youl;Chun, Dae-Sung;Eom, Mac;Kang, Yun-Suk;Park, Yong-Gul
    • Proceedings of the KSR Conference
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    • 2007.11a
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    • pp.517-528
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    • 2007
  • Domestic or international existing researches regarding rail damage factors are focused on laying, vehicle conditions, driving speed and driving habits and overlook characteristics of track structure (elasticity, maintenance etc). Also in ballast track, as there is no special lateral spring stiffness of track also called as ballast lateral resistance in concrete track, generally, existing study shows concrete track has 2 time shorter life cycle for rail replacement than ballast track due to abrasion. As a result of domestic concrete track design and operation performance review, concrete track elasticity is lower than track elasticity of ballast track resulting higher damage on rail and tracks. Generally, concrete track has advantage in track elasticity adjustment than ballast track and in case of Europe, in concrete track design, it is recommended to have same or higher performance range of vertical elastic stiffness of ballast track but domestically or internationally review on lateral spring stiffness of track is very minimal. Therefore, through analysis of service line track on site measurement and analysis on performance of maintenance, in this research, dynamic characteristic behaviors of commonly used ballast and concrete track are studied to infer elasticity of service line track and experimentally prove effects of track lateral spring stiffness that influence curved rail damage as well as correlation between track elasticity by track system and rail damage to propose importance of appropriate elastic stiffness level for concrete and ballast track.

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Detection of HCV-RNA by Reverse Transcription Polymerase Chain Reaction Using Biotinylated and Radioiodinated Primers (역전사 중합효소 연쇄반응(RT-PCR)에 의한 HCV-RNA의 검출 : Biotin 및 방사성옥소 표지 Primer로 구성된 Kit의 이용)

  • Ryu, Jin-Sook;Moon, Dae-Hyuk;Cheon, Jun-Hong;Chung, Yoon-Young;Park, Hung-Dong;Chung, Young-Hwa;Lee, Young-Sang
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.2
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    • pp.220-226
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    • 1994
  • This study was performed to evaluate the clinical applicability of the reverse transcription polymerase chain reaction (RT-PCR ) kit of HCV-RNA using biotinylated and radioiodinated primers. Study subjects were 118 patients with positive anti-HCV. HCV-RNA in patient's serum was extracted by guanidium thiocyanate method. After first amplification, the product was reamplified by primers labelled with biotin and I-125. The final amplification product was defected by counting the radioactivity after incubation in avidin coated tubes. In 51 samples, the test was repeated for evaluation of reproducibility. This new method was also compared with conventional RT-PCR methods in 34 samples from patients with chronic liver disease. The results were as follows ; 1) HCV-RNA was positive in 85(97%)of 88 patients with chronic liver disease, and in 23 (73%) of 30 patients with normal liver function. 2) In comparison with conventional method, HCV-RNA was detected in 32(94%) of 34 patients with new method, whereas in 27(79%) of the same group with conventional method. 3) Repeated test with new method in 52 samples demonstrated 82% of concordant result. In conclusion, new method with biotinylated and radioiodinated primers was more sensitive than conventional method. However, great care must be taken for quality control because there were considerable interassay variation and possiblity of false positivity and false negativity.

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Identification and Characterization of Lactobacillus salivarius subsp. salivarius from Korean Feces

  • Bae, Hyoung-Churl
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.05a
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    • pp.89-119
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    • 2004
  • This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

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Superoxide Dismutase Gene Expression in the Endotoxin-Treated Rat Lung (내독소에 의한 백서 폐장의 Superoxide Dismutase 유전자 발현에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.215-221
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    • 1994
  • Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

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Process Optimization of Ginseng Berry Extract Fermentation by Lactobacillus sp. Strain KYH isolated from Fermented Kimchi and Product Analysis (발효 김치로부터 분리한 Lactobacillus sp. Strain KYH를 이용한 진생베리 추출물 최적 발효 공정 확립 및 생성물의 특성 분석)

  • Ha, Yoo-Jin;Yoo, Sun-Kyun;Kim, Mee Ree
    • Journal of the East Asian Society of Dietary Life
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    • v.26 no.1
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    • pp.88-98
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    • 2016
  • The pharmacological effects of ginseng berry have been known to improve psychological function, immune activities, cardiovascular conditions, and certain cancers. It is also known that fermentation improves the bioavailability of human beneficial natural materials. Accordingly, we investigated the optimal fermentation conditions of ginseng berry extract with strain isolated from conventional foods. We also analyzed the fermentation product and its antioxidant activity. The bacterium isolated from fermented kimchi was identified as Lactobacillus sp. strain KYH. To optimize the process, fermentation was performed in a 5 L fermenter containing 3 L of ginseng berry extract at 200 rpm for 72 hr. Under optimized conditions, batch and fed-batch fermentations were performed. After fermentation, organic acids, amino acids, sugars, ginsenosides, and antioxidant activity were evaluated. The optimum fermentation conditions were determined as pH 7.0 and a temperature of $30^{\circ}C$, respectively. After fermentation, the amounts and compositions of organic acids, amino acids, sugars, ginsenosides, and antioxidant activity were altered. In comparing the distribution of ginsenosides with that before fermentation, the ginsenoside Re was a major product. However, amounts of ginsenosides Rb1, Rc, and Rd were reduced, whereas amounts of ginsenosides Rh1 and Rh2 increased. Total phenol content increased to 43.8%, whereas flavonoid content decreased to 19.8%. The DPPH radical scavenging activity and total antioxidant activity increased to 27.2 and 19.4%, respectively.