• Title/Summary/Keyword: RFLP.

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천연소재 활성 검토를 위한 간장해 동물모델에서의 장내세균 분석 요구

  • 김동구;송영재;소주련;강사행;박정향;고세웅;김동근;김태현;명제훈;진종식
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.120-120
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    • 2022
  • 장내세균의 구성과 각종 질환의 상관관계에 대한 연구가 활발히 수행되고 있다. 대장관련 질환 뿐 아니라, 당뇨, 비만 등 대사성 질환, 심지어 정신질환에까지 장내세균 구성의 영향이 미치는 것으로 밝혀졌다. 이에 다양한 천연 소재를 활용하여 각종 동물질환모델에서 생리활성을 검토할 때, 장내세균의 구성을 분석하여 그 상관관계를 밝힐 필요가 있다. 장내세균의 구성에 따라 질환의 양상이 달라질 수 있으며, 치료에 영향을 줄 수 있다. 동물실험을 수행할 때에도 실험동물 간 장내세균의 구성이 다르면 질환의 유도 또는 천연소재에 의한 치료 패턴이 달라질 수 있음을 가정하여 본 연구를 수행하였다. 본 연구에서는 EtOH와 LPS를 이용하여 간장해 동물모델을 구축하고, 장내세균 구성을 T-RFLP법으로 분석하여 그 상관관계를 검토하였고, tracheloside 등의 천연물을 투여함으로써 성분의 대사 패턴을 확인하였다. 이 결과는 천연소재의 간장해 개선 연구를 수행하기 위한 기초 결과로 활용될 수 있다.

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Spawning Patterns of Three Bitterling Fishes (Pisces: Acheilognathinae) in Relation to the Shell Size of Host Mussels (Unio douglasiae sinuolatus) (납자루아과(Pisces: Acheilognathinae) 담수어류 3종의 숙주조개(작은말조개; Unio douglasiae sinuolatus) 크기에 대한 산란양상)

  • Choi, Hee-kyu;Lee, Hyuk Je
    • Korean Journal of Environment and Ecology
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    • v.33 no.2
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    • pp.202-215
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    • 2019
  • This study was conducted to investigate the spawning preference of the Acheilognathinae fishes in relation to the shell size of host mussels after identifying the species of eggs and fries in the host mussel using our recently developed RFLP (Restriction Fragment Length Polymorphism) molecular marker at four sites [Hongcheon Naechoncheon (HN) and Deokchicheon (HD) from the North Han River basin and Jeongseon Goljicheon (JG) and Joyanggang (JJ) from the South Han River] in South Korea during May in each year between 2015 and 2018. The Acheilognathinae fish observed in the studied sites included one species (Acheilognathus signifer) in HN and JG, three species (Rhodeus uyekii, A. signifer, and Acheilognathus yamatsutae) in HD, and two species (A. signifer and Acheilognathus yamatsutae) in JJ, and we collected 982 host mussels (Unio douglasiae sinuolatus) that inhabited in all four sites. Using the RFLP molecular marker, we confirmed 46 eggs and fry of the Acheilognathinae fish (454 A. signifer, 43 Acheilognathus yamatsutae, and 149 Acheilognathus yamatsutae) in Unio douglasiae sinuolatus (N=163; 16.6%). We compare the average shell length, shell height, and shell width of mussels with [presence] eggs/fry and mussels without [absence] eggs/fry to examine the spawning preference according to the size of host mussels in each site. The results show that the shell length (1.98 mm), shell height (0.85 mm), and shell width (0.73 mm) of mussels with the eggs/fry were significantly larger (Mann-Whitney U test, P=0.002; difference=1.98 mm) than those of mussel without eggs/fry in HD where three species cohabitated. Although the shell length, shell height, and shell width of mussels with the eggs/fry were larger also in the other three sites, the difference was not statistically significant. In addition, we analyzed the mean number of spawned eggs and fry of each species and found $9.31{\pm}5.94$ R. uyekii, $2.86{\pm}2.45$ A.signifer, and $2.50{\pm}1.32$ A. yamatsutae. R. uyekii spawned 6.45-6.81 more eggs than A.signifer and A. yamatsutae on average per mussel, and it was statistically significant (Kruskal-Wallis test, P < 0.001). These findings indicate that the three species of Acheilognathinae fish tend to prefer larger mussels as their spawning hosts, and this tendency increases when the number of cohabitating bitterling fish species increases. Moreover, A.signifer and A. yamatsutae spawned a smaller number of eggs evenly in more host mussels while R. uyekii spawned many eggs on relatively fewer mussels. We found mussels (N=4) having the eggs/fry of two coexisting species, A. signifier and A. yamatsutae in HD and JJ where more than two bitterling fish species occurred. It suggests the interspecific competition taking place between the Acheilognathinae fishes for utilizing the same resource of mussels for spawning when two or more species cohabitate. This study is expected help to understand better the spawning patterns and reproductive ecology of the Acheilognathinae fishes, which will provide insightful information for advancing our understanding of their ecological relationships - mutualism or host-parasitism - with host mussels.

Polymorphisms of tumor necrosis factor-alpha promotor gene in Kawasaki disease and relation to the risk of coronary artery lesion (가와사키병에서의 TNF-alpha 유전자의 다형성 및 관상동맥 합병증과의 연관성)

  • Kim, Se-hwa;Yun, Jang-won;Lee, Young-hyuk;Cheon, Eun-jung
    • Clinical and Experimental Pediatrics
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    • v.52 no.4
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    • pp.476-480
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    • 2009
  • Purpose : The purpose of this study was to investigate the polymorphisms of the TNF-alpha promotor gene, its susceptibility to Kawasaki disease (KD) and to assess whether the TNF-alpha promotor gene polymorphism was related the risk of coronary artery lesions (CALs). Methods : From January 2003 to January 2007, 51 children (30 boys and 21 girls) with KD and 48 children forming an age-matched control group were studied. DNA from the peripheral blood of all the children was sampled, and the DNA polymorphisms of the 5' flanking regions of the TNF-alpha promoter gene at position -308 [guanine (G) to adenine (A)] were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Then, the relationship between KD and the TNF-alpha promotor gene polymorphisms was evaluated. Results : The A allele frequency of the -308 site of the TNF-alpha promotor gene was 17.6% (9/51) for children with KD and 6.8% (3/48) for the control group children, but this result was not statistically significant. Twenty-four patients experienced CALs within 60 days after the onset of symptoms. KD children with TNF-alpha -308 A allele had lower frequencies of CALs (12.5% versus 22.2%, P>0.05). Conclusion : The DNA polymorphism of the -308 site TNF-alpha gene was not associated with susceptibility to KD and a risk of CALs. Multicenter, large-scale randomized controlled trials are needed for further study.

New Evidences of Effect of Melanocortin-4 Receptor and Insulin-like Growth Factor 2 Genes on Fat Deposition and Carcass Traits in Different Pig Populations

  • Chen, J.F.;Xiong, Y.Z.;Zuo, B.;Zheng, R.;Li, F.E.;Lei, M.G.;Li, J.L.;Deng, C.Y.;Jiang, S.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1542-1547
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    • 2005
  • The Melanocortin-4 Receptor (MC4R) and Insulin-like Growth Factor 2 (IGF2) are two important candidate genes related to fat deposition and carcass traits. MC4R was found on study on human obesity and then was studied as candidate gene affecting food intake and fat deposition traits in mice and pigs. Insulin-like Growth Factor 2 (IGF2) gene plays an important role on tumor cell proliferation and muscle growth. It also affects fat traits and live weight in pigs. In this paper, MC4R and IGF2 were studied as two candidate genes associated with important economic traits such as fat deposition and carcass traits in five different pig populations. Taq I-PCR-RFLP and Bcn I-PCR-RFLP were respectively used to detect the polymorphism of genotypes of MC4R and IGF2 genes. Different MC4R genotype frequencies were observed in four populations. IGF2 genotype frequencies were also different in two populations. The results of association analysis show both MC4R and IGF2 genes were significantly associated with fat deposition and carcass traits in about 300 pigs. This work will add new evidence of MC4R and IGF2 affecting fat deposition and carcass traits in pigs and show that two genes can be used as important candidate genes for marker assistant selection (MAS) of growth and lean meat percentage in pigs.

Effects of a Porcine MC4R Polymorphism(892G>A) on Carcass Traits in Commercial Pigs (돼지 MC4R 유전자 892G>A 다형성이 비육돈의 도체형질에 미치는 영향)

  • Han, S.H.;Lee, S.S.;Ko, M.S.;Seong, P.N.;Park, B.Y.;Cho, I.C.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.569-576
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    • 2007
  • receptor(MC4R) gene and carcass traits was examined in randomly selected commercial pigs. A porcine MC4R gene was genotyped for Asp298Asn(nt. 892G>A) by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). A total of three genotypes, A/A, A/G, and G/G, were found with 28.8, 22.8, and 48.4% frequencies, respectively. In the whole population, pigs containing 892A/- showed significantly higher marbling score than those of homozygotes G/G(P<0.05). Two homozygotes, A/A and G/G showed lower in meat color score but higher in water holding capacity than those of heterozygotes A/G(P<0.01). However, the carcass weight of the barrows containing wild type -/G was significantly higher(i.e. more than 2.5kg) than those of homozygotes A/A(P<0.05). The effects of each genotype on carcass traits in the gilts were similar to those of the whole population, but not in barrows, suggesting an unknown sex-related effect on carcass traits. This study suggested that the genotype MC4R A/- could improve the meat quality in the commercial pig production. However, since the genetic polymorphism of MC4R gene differentially affected the carcass traits in sex-related manner, therefore, both parameters, the sex and genotype, should be considered for marker-assisted selection in commercial pig production.

QTL Mapping of Resistance to Gray Leaf Spot in Ryegrass: Consistency of QTL between Two Mapping Populations

  • Curley, J.;Chakraborty, N.;Chang, S.;Jung, G.
    • Asian Journal of Turfgrass Science
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    • v.22 no.1
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    • pp.85-100
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    • 2008
  • Gray leaf spot (GLS) is a serious fungal disease caused by Pyricularia oryzae Cavara, recently reported on the important turf and forage species, perennial ryegrass (Lolium perenneL.). This fungus also causes rice blast, which is usually controlled by host resistance, but durability of resistance is a problem. Few instances of GLS resistance have been reported in perennial ryegrass. However, two major QTL for GLS resistance have been detected on linkage groups 3 and 6 in an Italian x perennial ryegrass mapping population. To confirm that those QTL are still detectable in the next generation and can function in a different genetic background, a resistant segregant from this population has been crossed with an unrelated susceptible perennial clone, to form a new mapping population segregating for GLS resistance. QTL analysis has been performed in the new population, using two different ryegrass field isolates and RAPD, RFLP, and SSR marker-based linkage maps for each parent. Results indicate the previously identified QTL on linkage group 3 is still significant in the new population, with LOD and percent of phenotypic variance explained ranging from 2.0 to 3.5 and 5% to 10%, respectively. Also two QTL were detected in the susceptible parent, with similar LOD and phenotypic variance explained. Although the linkage group 6 QTL was not detected, the major QTL on linkage group 3 appears to beconfirmed. These results will add to our understanding of the genetic architecture of GLS resistance in ryegrass, which will facilitate its use in perennial ryegrass breeding programs.

Microbial community structures in the ground bed cultivation of Lentinula edodes using oak sawdust (참나무 톱밥을 이용한 표고 지면재배 과정의 주요 미생물 군집 분석)

  • Shin, Ji-Hye;Yun, Seo-Yeon;Nam, Ji-Hyun;Koo, Chang-Duck;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.221-230
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    • 2015
  • Ground bed cultivation method for shiitake mushroom (Phogo; Lentinula edodes) has been advanced to use the oak-sawdust composted by microorganisms. This method has some advantages in saving the labors and improving productivity. The aim of this study is to analyze microbial community in oak sawdust and to provide the information for efficient cultivation of shiitake mushroom. In process of composting, the proportions of thermophiles increased from 10% to 80%. This results show the microbial community succession from mesophilic bacteria to thermophilic bacteria. T-RFLP analysis and nucleotide sequencing of 16S rRNA gene demonstrated that the change of dominant bacteria coupled with shifts in each stages. The family of Enterobacteriaceae as a mesophilic bacteria were dominant (100%) in oak sawdust sample, but Amycolatopsis (49.0%) and Saccharopolyspora (26.5%) as a thermophilic Actinomycetes were dominant in the last day of composting. In hyphal growth stage, Leuconostoc (psychrophilic bacteria) was dominant (75.0%). The composting process of sawdust is very important for growth of mushroom mycelium. Therefore, there is need for the further study for the Amycolatopsis as thermophilic Actinomycetes.

Pests occurring on Cymbidium (심비디움에 발생하는 해충의 종류)

  • Cho, Myoung Rae;Jeon, Sung-Wook;Kang, Taek Joon;Kim, Hyung Hwan;Ahn, Seung-Joon;Yang, Chang Yeol
    • Korean journal of applied entomology
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    • v.52 no.4
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    • pp.403-408
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    • 2013
  • A survey of pest occurrence and status of farmer's pest management was conducted at 45 cymbidium farms in 10 major cultivation areas in Korea. The pest species collected from the cymbidium farms were identified as follows: Tetranychus urticae Koch, Frankliniella intonsa Trybom, Pinnaspis aspidistrae Signoret, Incilaria confusa Cockarel, Halyomorpha brevis Walker, Myzus persicae S$\ddot{u}$lzer, and Aphis gossypii Glover, Coccus hesperidum Linnaeus, Thrips flavus Schrank, and Thrips tabaci Lindeman. The two-spotted spider mite, T. urticae, was the key pest in cymbidium production, occurring on 45 farms, followed by scales (20 farms), slugs (6), thrips (8), aphids (5), and stinkbug (1). PCR-RFLP of the rDNA ITS2 region revealed that two thrips species, Thrips flavus Schrank and Thrips tabaci Lindeman, occur on cymbidium farms. Therefore, it is necessary for the cymbidium farmers to establish an integrated pest management system to meet quarantine standards.

Characterization of Phosphoinositide-3-kinase, Class 3 (PIK3C3) Gene and Association Tests with Quantitative Traits in Pigs

  • Kim, J.H.;Choi, B.H.;Lim, H.T.;Park, E.W.;Lee, S.H.;Seo, B.Y.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1701-1707
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    • 2005
  • This study deals with the characterization of porcine PIK3C3 and association tests with quantitative traits. PIK3C3 belongs to the class 3 PI3Ks that participate in the regulation of hepatic glucose output, glycogen synthase, and antilipolysis in typical insulin target cells such as those in the such as liver, muscle system, and fat. On the analysis of full-length mRNA sequence, the length of the PIK3C3 CDS was recorded as 2,664 bps. As well, nucleotide and amino acid identities between human and pig subjects were 92% and 99%, respectively. Five SNPs were detected over 5 exons. We performed genotyping by using a SNP C2604T on exon24 for 145 F$_2$ animals (from a cross between Korean native boars and Landrace sows) by PCR-RFLP analysis with Hpy8I used to investigate the relationship between growth and fat depot traits. In the total association analysis, which doesn' consider transmission disequilibrium, the SNP showed a significant effect (p<0.05) on body weight and carcass fat at 30 weeks of age as well as a highly significant effect (p<0.01) on back fat. In an additional sib-pair analysis, C allele still showed positive and significant effects (p<0.05) on back fat thickness and carcass fat. Moreover, the effects of C allele on the means of within-family components for carcass fat and back fat were estimated as 2.76 kg and 5.07 mm, respectively. As a result, the SNP of porcine PIK3C3 discovered in this study could be utilized as a possible genetic marker for the selection of pigs that possess low levels of back fat and carcass fat at the slaughter weight.

Analysis of Populus cpDNA by Restriction Fragment Length Polymorphism(RFLP) Technique (RFLP기법(技法)을 이용(利用)한 포플러 엽록체(葉綠體) DNA의 분석(分析))

  • Lee, J.S.;Noh, E.W.;Lee, S.K.;Kwon, K.W.
    • Journal of Korean Society of Forest Science
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    • v.83 no.1
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    • pp.20-24
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    • 1994
  • In woody species with a long life span, the studies on inheritance of any trait may be very time consuming and laborious. Chloroplast DNA(cpDNA) has been a valuable tool in such studies since it has several unique features such as limited genome size and cytoplasmic inheritance. In the present study, cpDNAs from five different species of Populus(P. alba, P. glandulosa, P. alba${\times}$P. glandulosa, P. davidiana, and P. nigra), and Nicotiana tabacum were compared with regard to restriction fragment length polymophism. The results showed that cpDNAs among the species were very conserved, although some polymorphisms were observed when the DNAs were digested with restriction enzyme EcoRI or KphI. The other enzymes (Bgl II, and PstI) tested produced identical restriction fragmentation pattern among the species. However, cpDNAs from all the five Populus species showed different restriction fragmentation pattern from that of tobacco with the four restriction enzymes tested. Southern hybridization with tobacco rbcL gene fragment as a probe also produced identical pattern among Populus species. The results indicate that cpDNAs in the genus are very well conserved during evolution.

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