• Title/Summary/Keyword: gene gun

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Genetic Analysis of Pod Dehiscence in Soybean

  • Kang Sung Taeg;Kim Hyeun Kyeung;Baek In Youl;Chung Moung Gun;Han Won Young;Shin Doo Chull;Lee Suk-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.4
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    • pp.281-285
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    • 2005
  • Pod dehiscence (PD), defined as the opening of pods along both the dorsal and ventral sutures, causes the seed to shatter in the field before harvesting and results in loss of seed yields. However, breeding for resistance to PD is difficult due to the complicated genetic behavior and environmental interaction. The objective of the present research was to analyze the genetic behavior of PD for improving the breeding efficiency of resistance to PD in soybean. PD after oven-drying the sampled pod at $40^{\circ}C$ for 24 hours was the most reliable to predict the degree of PD tested in the field. Keunolkong, a dehiscent parent, was crossed with non-dehiscent parents, Sinpaldalkong and Iksan 10. Using their $F_1\;and\;F_2$ seeds, PD was measured after oven drying the pod at $40^{\circ}C$ for 24 hours. The gene conferring PD behaved in different manners depending on the genetic populations. In the Keunolkong$\times$Sinpaldalkong population, PD seemed to be governed by single major recessive gene and minor genes, while several genes were probably involved in the resistance to pod dehiscence in the Keunolkong$\times$Iksan 10 population. Heritability for PD estimated in F2 population showed over $90\%$ in the two populations. High heritability of PD indicated that selection for resistance to PD should be effective in a breeding program. In addition, genetic mapping of quantitative locus (QTL) for PD in both populations may reveal that genes conferring PD are population-specific.

Identification of the Vibrio vulnificus fexA Gene and Evaluation of its Influence on Virulence

  • JU HYUN-MOK;HWANG IN-GYUN;WOO GUN-JO;KIM TAE SUNG;CHOI SANG HO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1337-1345
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    • 2005
  • Vibrio vulnificus is the causative agent of foodborne diseases such as gastroenteritis and life-threatening septicemia. Microbial pathogenicity is a complex phenomenon in which expression of numerous virulence factors is frequently controlled by a common regulatory system. In the present study, a mutant exhibiting decreased cytotoxic activity toward intestinal epithelial cells was screened from a library of V. vulnificus mutants constructed by a random transposon mutagenesis. By a transposon-tagging method, an open reading frame, fexA, a homologue of Escherichia coli areA, was identified and cloned. The nucleotide and deduced amino acid sequences of the fexA were analyzed, and the amino acid sequence of FexA from V. vulnificus was $84\%\;to\;97\%$ similar to those of AreA, an aerobic respiration control global regulator, from other Enterobacteriaceae. Functions of the FexA were assessed by the construction of an isogenic mutant, whose fexA gene was inactivated by allelic exchanges, and by evaluating its phenotype changes in vitro and in mice. The disruption of fexA resulted in a significant alteration in growth rate under aerobic as well as anaerobic conditions. When compared to the wild-type, the fexA mutant exhibited a substantial decrease in motility and cytotoxicity toward intestinal epithelial cell lines in vitro. Furthermore, the intraperitoneal $LD_{50}$ of the fexA mutant was approximately $10^{1}-10^{2}$ times higher than that of parental wild-type. Therefore, it appears that FexA is a novel global regulator controlling numerous genes and contributing to the pathogenesis as well as growth of V. vulnificus.

Detection of Human Cytomegalovirus UL97 D605E Mutation in Korean Stem Cell Transplantation Recipients and Donors

  • Lee, Gyu-Cheol;Choi, Su-Mi;Lee, Chan Hee;Lee, Dong-Gun;Choi, Jung-Hyun;Yoo, Jin-Hong
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1154-1158
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    • 2013
  • Ganciclovir resistance of human cytomegalovirus is associated with mutations in the viral UL97 gene and poses severe problems for immunocompromised patients. In this study, PCR-based restriction fragment length polymorphism and sequencing analyses detected the UL97 D605E mutation in all five clinical isolates from patients with ganciclovir-resistant human cytomegalovirus infection during prolonged ganciclovir therapy, whereas the M460V mutation was only present in 1 of 5 isolates. On the other hand, the detection rates of the D605E mutation in the stored available DNA samples from the donor and allogeneic stem cell transplantation recipients were 66.7% and 93.7%, respectively, suggesting that the presence of D605E mutation was not associated with the ganciclovir exposure. Although the D605E mutation may not be related to ganciclovir resistance, we suggest that this mutation could be an important molecular marker of human cytomegalovirus evolution in East Asian countries. Moreover, the restriction fragment length polymorphism method using the restriction enzyme HaeIII, which is generally used to detect the UL97 A591V mutation, could also detect the D605E mutation and may therefore be a useful tool for future research on the investigation of UL97 gene mutations.

Use of 16S-23S rRNA Intergenic Spacer Region for Species-specific Primer Developed of Vibrio Ichthyoenteri (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio ichthyoenteri Species-specific Primer 개발)

  • Moon Young-Gun;Heo Moon-Soo
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.117-124
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    • 2005
  • Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

Species-specific variation of RPA-interacting protein (RIP) splice isoforms

  • Kim, Kwang-Soo;Lee, Eun-Ju;Lee, Seung-Hoon;Seo, Tae-Gun;Jang, Ik-Soon;Park, Jun-Soo;Lee, Je-Ho
    • BMB Reports
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    • v.42 no.1
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    • pp.22-27
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    • 2009
  • Replication Protein A (RPA) is a single stranded DNA-binding protein involved in DNA metabolic activities such as replication, repair, and recombination. RPA-Interacting Protein $\alpha$ ($RIP{\alpha}$) was originally identified as a nuclear transporter of RPA in Xenopus. The human $RIP{\alpha}$ gene encodes several splice isoforms, of which $hRIP{\alpha}$ and $hRIP{\beta}$ are the major translation products in vivo. However, limited information is available about the alternative splicing of $RIP{\alpha}$ in eukaryotes, apart from that in humans. In this study, we examined the alternative splicing of RIP{\alpha} in the Drosophila, Xenopus, and mouse system. We showed that the number of splice isoforms of RIP{\alpha} was species-specific, and displayed a tendency to increase in higher eukaryotes. Moreover, a mouse ortholog of $hRIP{\alpha}$, $mRIP{\beta}2$, was not SUMOylated, in contrast to $hRIP{\alpha}$. Based on these results, we suggest that the $RIP{\alpha}$ gene gains more splice isoforms and additional modifications after molecular evolution.

Quantitative distribution of denitrifying bacteria with nirS and nirK in MLE and A/O process (MLE와 A/O 공정에서의 nirS 와 nirK 를 가진 탈질미생물의 정량적 분포)

  • Lim, Dong-Seok;Kim, Yun-Jung;Kim, Hyung-Gun;Park, Seung-Guk;Chung, Tai-Hak
    • Journal of Korean Society of Water and Wastewater
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    • v.26 no.4
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    • pp.591-598
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    • 2012
  • Denitrification is an important biological mechanism in wastewater treatment process because this process is technically to remove nitrogen from water to air. There have been lots of study about denitrification engineering and molecular biological research about denitrifying bacteria, respectively. However, combination of these researches was unusual and rare. This study is about the correlation between quantity of denitrifying bacteria and denitrification potential, and consists of NUR batch test as analysis method of denitrification potential and quantitative molecular analysis for denitrifying bacteria. Three reactors (A/O, MLE and A/O of nitrogen deficiency) are operated to get activated sludge with various denitrification potential. All samples which were acquired from reactors were measured denitrification potential by NUR test and NUiR test. Also, Real-time PCR was conducted for quantification of denitrifying bacteria composition in activated sludge. The various denitrification potentials were measured in the reactors. The denitrifiaction potential was the highest in MLE process and the reactor of the nitrogen deficiency showed the lowest. Genomic DNA of activated sludge was obtained and consequently, real-time PCRuse the primer sets of nirK and nirS were conducted to quantify genes involving denitrification reductase production. As the result of real-time PCR, nirK gene showed more significant influence on denitrification potential comapred with nirS gene.

Screening of Anti-cancer Compounds Originated from Filamentous Fungi (Monascus sp.) (사상성 곰팡이 (Monascus sp.) 유래 항암 물질의 탐색)

  • Sin, Yeong-Min;Park, Hae-Ryoun;An, Won-Gun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.3
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    • pp.671-676
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    • 2005
  • In this study, we investigated the antioxidant effect of extract from Monascus pillosus, on the human wild-type p53 and p21 expressing A549 lung epithelial cell line and MCF-7 mammary adenocarcinoma cell line stimulated by NO. $P21^{waf/cip1}$ was identified as a gene induced in senescent cells. It is a cyclin-dependent kinase inhibitor and has been shown to cause cell cycle arrest and apoptosis. While p53-regulated stimulation of p21 appears to be central for the permanent growth-arrest, the role of p21 in p53-triggered cell death is unclear. Low dose of sodium nitroprusside (SNP) induced the development of senescence associated with increased expression of p53 and p21 in A549 cells. Inhibition of p21 transactivating activity requires high level correlates with the amount of p53 necessary to cause cell death. Association of p21 and p53 results in inhibition of p21-stimulated transcription. This requires a higher p53 level than is necessary for transcriptional activation of endogenous p53-responsive gene but correlates well with the level of p53 necessary to cause cell death. Exposure to W-1 inhibited oxidative stresses-induced senescence-like arrest, resulting in a significant reduction in p53 and p21 steady state levels. These results suggest that p53 and p21 play a central role in the onset of senescence. Thus, it is important to emphasize control of oxidative balance in tumor prevention and aging.

Ramlibacter ginsenosidimutans sp. nov., with Ginsenoside-Converting Activity

  • Wang, Liang;An, Dong-Shan;Kim, Song-Gun;Jin, Feng-Xie;Kim, Sun-Chang;Lee, Sung-Taik;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.311-315
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    • 2012
  • A novel ${\beta}$-proteobacterium, designated BXN5-$27^T$, was isolated from soil of a ginseng field of Baekdu Mountain in China, and was characterized using a polyphasic approach. The strain was Gram-staining-negative, aerobic, motile, non-spore-forming, and rod shaped. Strain BXN5-$27^T$ exhibited ${\beta}$-glucosidase activity that was responsible for its ability to transform ginsenoside $Rb_1$ (one of the dominant active components of ginseng) to compound Rd. Phylogenetic analysis based on 16S rRNA gene sequences showed that this strain belonged to the family Comamonadaceae; it was most closely related to Ramlibacter henchirensis $TMB834^T$ and Ramlibacter tataouinensis$TTB310^T$ (96.4% and 96.3% similarity, respectively). The G+C content of the genomic DNA was 68.1%. The major menaquinone was Q-8. The major fatty acids were $C_{16:0}$, summed feature 4 (comprising $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2OH), and $C_{17:0}$ cyclo. Genomic and chemotaxonomic data supported the affiliation of strain BXN5-$27^T$ to the genus Ramlibacter. However, physiological and biochemical tests differentiated it phenotypically from the other established species of Ramlibacter. Therefore, the isolate represents a novel species, for which the name Ramlibacter ginsenosidimutans sp. nov. is proposed, with the type strain being BXN5-$27^T$ (=DSM $23480^T$ = LMG $24525^T$ = KCTC $22276^T$).

Effects of Vitamins E and C on Human BreastCancer Cell Growth in the Presence of Various Fatty Acids

  • Kim, Gun-Hee;Cho, Il-Jin;Oh, Sun-Hee;Park, Hee-Sung;Cho, Sung-Hee
    • Preventive Nutrition and Food Science
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    • v.3 no.1
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    • pp.85-91
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    • 1998
  • To investigate the effects of antioxidative vitamins in combination with various fatty acids on breast cancer cell proliferation, MDA-MB231 human breast cancer cells were cultured for 3 days in the serum-free Iscove's modified Dulbecco's medium (IMDM) supplemented with 1.25mg/ml delipidized bovine serum albumin and 10㎍/ml insulin. Alpha-tocopherol, ascorbic acid or both vitamins were added to the medium at the concentrations of 10 and 50μM in the presence of 3μg/ml of oletic(Oa), linoleic(LA) α-linoleinic(LNA) and docosahexaenoic acid(DHA). Cell growth was reduced significantly by α-tocopherol in a dose-dependent manner, but not affected by ascorbic aicd. The four different fatty acids did not have significant effects on cell growth, although DHA exerted inhibitory effect on the growth after 1 day. However, the each fatty acid was well incorporated into celluar lipid as such or elongated forms. Addition of α-tocopherol remarkably increased its celluar contents and reduced cellular levels of thiobarbituric acid substances (TBARS) that were elevated notably in the presence of DHA in the culture media. But ascorbic acid addition did not change much of either cellular α-tocopherol or TBARS contents. northern blot hybridization showed that tumor supressor gene ρ53 was most highly expressed by the combination of ρ-tocopherol and DHA in 8 hours of cell culture. In conclusion , the growth inhibitory effect of vitamin E suggests that breast cancer cell proliferation is reduced by the mechanism other than cytotoxicity of lipid peroxide and it is related to expressionof tumor supprosser gene p53, that can be increased by both vitamin E and n-3 fatty acid, DHA.

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Evaluation of the microbiome composition in particulate matter inside and outside of pig houses

  • Hong, Se-Woon;Park, Jinseon;Jeong, Hanna;Kim, Minseok
    • Journal of Animal Science and Technology
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    • v.63 no.3
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    • pp.640-650
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    • 2021
  • Particulate matter (PM) produced in pig houses may contain microbes which can spread by airborne transmission, and PM and microbes in PM adversely affect human and animal health. To investigate the microbiome in PM from pig houses, nine PM samples were collected in summer 2020 inside and outside of pig houses located in Jangseong-gun, Jeollanam-do Province, Korea, comprising three PM samples from within a nursery pig house (I-NPH), three samples from within a finishing pig house (I-FPH), and three samples from outside of the pig houses (O-PH). Microbiomes were analyzed using 16S rRNA gene amplicon sequencing. Firmicutes was the most dominant phylum and accounted for 64.8%-97.5% of total sequences in all the samples, followed by Proteobacteria (1.4%-21.8%) and Bacteroidetes (0.3%-13.7%). In total, 31 genera were represented by > 0.3% of all sequences, and only Lactobacillus, Turicibacter, and Aerococcus differed significantly among the three PM sample types. All three genera were more abundant in the I-FPH samples than in the O-PH samples. Alpha diversity indices did not differ significantly among the three PM types, and a principal coordinate analysis suggested that overall microbial communities were similar across PM types. The concentration of PM did not significantly differ among the three PM types, and no significant correlation of PM concentration with the abundance of any potential pathogen was observed. The present study demonstrates that microbial composition in PM inside and outside of pig houses is similar, indicating that most microbe-containing PM inside pig houses leaks to the outside from where it, along with microbe-containing PM on the outside, may re-enter the pig houses. Our results may provide useful insights regarding strategies to mitigate potential risk associated with pig farming PM and pathogens in PM.