• 제목/요약/키워드: Chromosome analysis

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Effect of Butyrate on Adenovirus-Mediated Herpes Simplex Virus Thymidine Kinase Gene Therapy (Butyrate가 Adenoviral Vector로 이입한 Herpes Simplex Virus Thymidine Kinase 유전자치료에 미치는 영향)

  • Park, Jae-Yong;Kim, Jeong-Ran;Chang, Hee-Jin;Kim, Chang-Ho;Park, Jae-Ho;Jung, Tae-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.3
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    • pp.587-595
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    • 1998
  • Background: Recombinant adenovirus hold promise as vectors to carry therapeutic genes for several reasons: 1) they can infect both dividing and non-dividing cells; 2) they have the ability to directly transduce tissues in vivo; 3) they can easily be produced in high titer; and 4) they have an established record of safety as vaccination material. However, one of the major limitation in the use of adenoviruses is that transgene expression is quite short because adenovirusees insert their DNA genome episomally rather than by chromosomal integration, and an immune response against the virus destroys cells expressing the therapeutic gene. Since sodium butyrate has been reported to induce adenovirus-mediated gene expression, we hypothesized that treatment of tumor cells, transduced with herpes simples virus thymidine kinase(HSVtk) gene using adenoviral vector, with butyrate could augment the effect of gene therapy. Methods: We transduced HSVtk gene, driven by the cytomegalovirus promoter, into REN cell line(human mesothelioma cell line). Before proceeding with the comparison of HSVtk/ganciclovir mediated bystander killing, we evaluated the effect of butyrate on the growth of tumor cells in order to rule out a potential antitumor effect of butyrate alone, and also on expression of HSVtk gene by Western blot analysis. Then we determined the effects of butyrate on bystander-mediated cell killing in vitro. Results: There was no inhibition of growth of cells exposed to butyrate for 24 hours at a concentration of 1.5mM/L. Toxic effects were seen when the concentration of butyrate was greater than 2.0mM/L. Gene expression was more stable and bystander effect was augmented by butyrate treatment of a concentration of 1.5mM/L. Conclusion: These results provide evidence that butyrate can augment the efficiency of cell killing with HSVtk/GCV system by inducing transgene expression and may thus by a promising new approach to improve responses in gene therapy using adenoviral vectors.

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Association of SNPs from iNOS and TLR-4 Genes with Economic Trait in Chicken (닭의 iNOS와 TLR-4 유전자 내 변이와 경제 형질 간의 연관성 분석)

  • Lim, Hee Kyong;Han, Jung-Min;Oh, Jae Don;Lee, Hak Kyo;Jeon, Gwang Joo;Lee, Jun Heon;Seo, Dong Won;Cahyadi, Muhammad;Song, Ki Duk;Choi, Kang Duk;Kong, Hong Sik
    • Korean Journal of Poultry Science
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    • v.40 no.2
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    • pp.83-89
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    • 2013
  • iNOS (Inducible nitric oxide synthase) and TLR-4 (Toll-like Receptor-4) play crucial roles in innate immunity of poultry. iNOS has been mapped to chicken chromosome 14 and implicated in a variety of chicken diseases. iNOS possesses potent antimicrobial activity, including the inhibition of microbes replication in vitro. TLR-4 is a pathogen associated molecular-pattern receptor for bacterial product, such as LPS (lipopolysaccharides) found in Gram negative bacteria, that triggers pro-inflammatory cytokine expression after engagement with ligands. In the previous studies, genetic analysis of iNOS and TLR-4 revealed the possible association of mutation in these genes with the intestinal microflora of cecum when infected with Salmonella spp. This study was aimed to augment previous findings, which show the association of iNOS (C14513T) and TLR-4 (G4409T) polymorphisms with economic traits in Korean Native Black (KNB), Rhode Island Red (RIR) and Cornish chickens. Investigation in the effect of SNPs on economic traits (layday, layw, layno, bw150, bw270, layw270) was conducted. iNOS (C14513T) had a significant effect on the average body weight at 270 days of age (p<0.05) in both KNB and RIR, whereas TLR-4 (G4409T) showed no significant correlation with all traits (p>0.05). The results obtained from using the candidate genes can be useful for the genetic improvement of body weight in both KNB and RIR breeds.

Analysis of Quantitative Trait Loci (QTLs) for Seed Size and Fatty Acid Composition Using Recombinant Inbred Lines in Soybean (콩 재조합자식계통을 이용한 콩 종자의 크기와 지방산 조성의 양적 형질 유전자좌 분석)

  • Kim, Hyeun-Kyeung;Kim, Yong-Chul;Kim, Sun-Tae;Son, Beung-Gu;Choi, Yong-Whan;Kang, Jum-Soon;Park, Young-Hoon;Cho, Young-Son;Choi, In-Soo
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1186-1192
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    • 2010
  • Soybean [Glycine max(L.) Merr.] is an important crop, accounting for 48% of the world market in oil crops. Improvements in economic traits, such as quality and oil constituents, arethe most important objectives in soybean breeding. The objective of this study was to identify quantitative trait loci (QTLs) that control seed size and fatty acid contents in soybean. 115 $F_{2:10}$ recombinant inbred lines (RIL) developed from a cross of 'Keunolkong' and 'Iksan10' were used. Narrow-sense heritability estimates based on a plot mean on 100 seed weight, saturated fatty acid (palmitic acid + stearic acid), and oleic, linoleic, and linolenic acid content were 0.72, 0.60, 0.83, 0.77 and 0.81, respectively. The 100 seeds weight was related to seven QTLs located on chromosomes 1, 3, 8, 9, 16 and 17. Two independent QTLs for saturated fatty acid content were identified on chromosomes 17 and 19. Five independent QTLs for oleic acid content wereidentified on chromosomes7, 11, 14, 16 and 19. Five QTLs for linoleic acid content were located on chromosomes 2, 11, 14, 16 and 19. Three QTLs for linolenic acid content were located on chromosomes 8, 10 and 19. Oleic, linoleic, and linolenic acid had one major common QTL on chromosome 19. Thus, linoleic and linolenic acid content were identified as common QTLs.

Analysis of Telomere Length and Telomerase Activity of Tissues in Korean Native Chicken (한국 재래닭의 주령별 각 조직의 텔로미어 함량과 텔로머레이스 활성도 분석)

  • Jung G.S.;Cho E.J.;Choi D.S.;Lee M.J.;Park C.;Jeon I.S.;Sohn S.H.
    • Korean Journal of Poultry Science
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    • v.33 no.2
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    • pp.97-103
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    • 2006
  • Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Even though telomere length and telomerase activity have been studied extensively, very little is known to analyze the telomere dynamics in chicken cells. This study was carried out to analyze the telomere distribution and telomerase activity of Korean Native Chicken cells along with aging. The cells were collected from brain, heart, liver, kidney and germinal tissues during physiological stages. Telomere distribution was analyzed by Quantitative-Fluorescence in situ Hybridization (Q-FISH) techniques using the chicken telomeric DNA probe. Telomerase activity was performed by Telomeric Repeat Amplification Protocol (TRAP) assay. In results, the telomeres of chicken were found at the ends of all chromosomes with the interstitial telomeres on chromosomes 1, 2 and 3. The amount of telomeres on chicken cells was decreased along with aging in most tissues. Furthermore, the telomere quantity was significantly different among tissues. The relative amount of telomeres in proliferous cells such as testis cells had much more than those of liver, brain, heart, blood and kidney cells. The telomerase activity was down-regulated in cells of brain, heart and liver tissues. Whereas gonadal cells showed a constitutive activity of telomerase during all stage of life. In conclusions, the telomere quantity and telomerase activity in chicken are closely relate to cell proliferation and tissue specificity during developmental stages and aging. There is also closely correlated between the amounts of telomeric DNA and telomerase activity in chicken tissues.

Decitabine in the Treatment of Acute Myeloid Leukemia and Myelodysplastic Syndromes, Which Combined with Complex Karyotype Respectively

  • Gao, Su;Li, Zheng;Fu, Jian-Hong;Hu, Xiao-Hui;Xu, Yang;Jin, Zheng-Ming;Tang, Xiao-Wen;Han, Yue;Chen, Su-Ning;Sun, Ai-Ning;Wu, De-Pei;Qiu, Hui-Ying
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6627-6632
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    • 2015
  • Background: We conducted a study exploring the clinical safety and efficacy of decitabine in patients with acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS), combined with a complex karyotype. Materials and Methods: From April 2009 to September 2013, a total of 35 patients with AML/MDS combined with a complex karyotype diagnosed in the First Affiliated Hospital of Soochow University were included for retrospective analysis. All patients were treated with decitabine alone ($20mg/m^2$ daily for 5 days) or combination AAG chemotherapy (Acla 20mg qod*4d, Ara-C $10mg/m^2$ q12h*7d, G-CSF $300{\mu}g$ qd, the dose of G-CSF adjusted to the amount in blood routinely). Results: In 35 patients, 15 exhibited a complete response (CR), and 6 a partial response (PR), the overall response rate (CR+PR) being 60% (21 of 35). Median disease-free survival was 18 months and overall survival was 14 months. In the 15 MDS patients with a complex karyotype, the CR rate was 53.3% (8 of 15); in 20 AML patients with complex karyotype, the overall response rate was 65% (13 of 20). The response rate of decitabine alone (22 cases) was 56.5% (13 of 22), while in the combination chemotherapy group (13 cases), the effective rate was 61.5% (8 of 13)(P>0.05). There are 15 patients with chromosome 7 aberration, after treatment with decitabine, 7 CR, 3 PR, overall response rate was 66.7% (10 of 15). Of 18 patients with 3 to 5 kinds of chromosomal abnormalities, 66.7% demonstrated a response; of 17 with more than 5 chromosomal abnormalities, 52.9% had a response. In the total of 35 patients, with one course (23 patients) and ${\geq}$two courses (12 patients), the overall response rate was 40.9% and 92.3% (P<0.05). Grade III to IV hematological toxicity was observed in 27 cases (75%). Grade III to IV infections were clinically documented in 7 (20%). Grades I to II non-hematological toxicity were infections (18 patients), haematuria (2 patients), and bleeding (3 patients). With follow-up until September 2013, 7 patients were surviving, 18 had died and 10 were lost to follow-up. In the 6 cases who underwent allogeneic hematopoietic stem cell transplantation (HSCT) all were still relapse-free survivors. Conclusions: Decitabine alone or combination with AAG can improve outcome of AML/MDS with a complex karyotype, there being no significant difference decitabine in inducing remission rates in patients with different karyotype. Increasing the number of courses can improve efficiency. This approach with fewer treatment side effects in patients with a better tolerance should be employed in order to create an improved subsequent chance for HSCT.

Evaluation of Germplasm and Development of SSR Markers for Marker-assisted Backcross in Tomato (분자마커 이용 여교잡 육종을 위한 토마토 유전자원 평가 및 SSR 마커 개발)

  • Hwang, Ji-Hyun;Kim, Hyuk-Jun;Chae, Young;Choi, Hak-Soon;Kim, Myung-Kwon;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.30 no.5
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    • pp.557-567
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    • 2012
  • This study was conducted to achieve basal information for the development of tomato cultivars with disease resistances through marker-assisted backcross (MAB). Ten inbred lines with TYLCV, late blight, bacterial wilt, or powdery mildew resistance and four adapted inbred lines with superior horticultural traits were collected, which can be useful as the donor parents and recurrent parents in MAB, respectively. Inbred lines collected were evaluated by molecular markers and bioassay for confirming their disease resistances. To develop DNA markers for selecting recurrent parent genome (background selection) in MAB, a total of 108 simple sequence repeat (SSR) primer sets (nine per chromosome at average) were selected from the tomato reference genetic maps posted on SOL Genomics Network. Genetic similarity and relationships among the inbred lines were assessed using a total of 303 polymorphic SSR markers. Similarity coefficient ranged from 0.33 to 0.80; the highest similarity coefficient (0.80) was found between bacterial wilt-resistant donor lines '10BA333' and '10BA424', and the lowest (0.33) between a late blight resistant-wild species L3708 (S. pimpinelliforium L.) and '10BA424'. UPGMA analysis grouped the inbred lines into three clusters based on the similarity coefficient 0.58. Most of the donor lines of the same resistance were closely related, indicating the possibility that these lines were developed using a common resistance source. Parent combinations (donor parent ${\times}$ recurrent parent) showing appropriate levels of genetic distance and SSR marker polymorphism for MAB were selected based on the dendrogram. These combinations included 'TYR1' ${\times}$ 'RPL1' for TYLCV, '10BA333' or '10BA424' ${\times}$ 'RPL2' for bacterial wilt, and 'KNU12' ${\times}$ 'AV107-4' or 'RPL2' for powdery mildew. For late blight, the wild species resistant line 'L3708' was distantly related to all recurrent parental lines, and a suitable parent combination for MAB was 'L3708' ${\times}$ 'AV107-4', which showed a similarity coefficient of 0.41 and 45 polymorphic SSR markers.

Linkage Analysis of the Resistance Genes to Whitebacked Planthopper (Sogatella furcifera Horvath) in Rice (수도의 흰등멸구(Sogatella furcifera Horvath)에 대한 저항성 유전자 연관분석)

  • ;Mun-Hue Heu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.29 no.2
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    • pp.136-151
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    • 1984
  • The purpose of this study is to find out the linkage relationship of the resistance genes Wbph1 and Wbph2 which are known to be present in the rice cultivar N22 and ARC 10239 respectively, with the genetic markers which are identified as the specific linkage tester. Crosses were made between the resistant parents and the genetic marker stocks and their F$_2$ populations were grown out in the field. The genetic segregations of the marker character were studied and the seeds were harvested individual plant base. These F$_3$ seeds were grown into plant-line base in the greenhouse and their responses to the whitebacked planthopper were tested. Then the linkage relationship between the F$_2$ plant marker character and the F$_3$ resistance responses to the whitebacked planthopper were examined. In the F$_2$ generation of the crosses between the resistant parent N22 and the genetic marker stocks, the genetic markers, such as lg, d-t, g, la, bl and gl, showed the segregation of 3 dominance to 1 recessiveness, and the Bh marker segregated into 9:7 ratio. Another 4 marker genes, such as Cl, gh, Lh and bc, did not show the good fittness to the expected value. In the F$_2$ generation of the crosses between the resistant parent ARC 10239 and the genetic marker stocks, the genetic markers, such as Cl, lg, Pn, g, la, bl and gl, showed the segregation of 3 dominance to 1 recessiveness, and the Bh gene segregation fitted well to the 9:7. The rest 4 genetic markers, such as gh, Lh, nl and be, did not show the good fitness to the expected ratio. The resistance genes Wbphl of N22 and the Wbph2 of ARC 10239 appeared to be single dominant gene each. The Wbphl gene was linked with the marker gene, liguleless (lg) of linkage group II with the recombination value of 36.8%, and with the black hull (Bh) with the value of 35.9%. The Wbph2 gene appeared to be independent of all the markers tested here, such as Cl, lg, Pn, g, Lh, la, nl, bl, bc, gl, Bh, of linkage gtoup I, II, III, IV, VI, VII, VIII, IX, X, XI, and XII respectively. That the Wbph2 linkage relations were not investigated was regarded as the causes that the tested marker genes on the chromosome were located with the resistance gene at the distant loci, and of the phenctypic properties of the marker characters. The Wbph2 linkage relations should be reexamined in the cross combinations of linkage group Ⅶ, Ⅷ, Ⅹ and XII including linkage group V which was not tested in this experiment.

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Genetic diversity of Clostridium perfringens form food-poisoning outbreak in Gyeonggi-do, 2013-2014 (경기도 식중독에서 분리된 Clostridium perfringens의 유전적 특성 분석)

  • Park, Sung-Hee;Choi, Ok-Kyung;Jeong, Jin-A;Kim, Woon-Ho;Lee, Yea-Eun;Park, Kwang-Hee;Yoon, Mi-Hye
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.286-297
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    • 2016
  • Clostridium perfringens is both a ubiquitous environmental bacterium and a major cause of human gastrointestinal disease, and C. perfringens food poisoning ranks among the most common gastrointestinal diseases in developed countries. 120 isolates of C. perfringens were obtained from food-poisoning outbreaks in 2013~2014, Gyeonggi-do. Using PCR, all 120 isolates were identified as C. perfringens type A. Of the tested isolates, 49 isolates carried the cpe gene, 71 isolates carried the cpb2 gene. The outbreak cases of cpb2 and cpe /cpb2 genes were 7 and 7, whereas the outbreak cases of cpe-gene were 2. The epidemiological relationship between C. perfringens isolates has previously been investigated chiefly by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). The genetics relatedness of the isolates raged from 53.5-100% and 75 district PFGE type were observed. The PFGE results revealed a wide genetic diversity among the 64 cpb2 carrying isolates (except 7 isolates), while 46 cpe-carrying isolates (except 3 isolates) showed a high genetic similarity. The MLST analysis revealed that 14 cpe isolates (cpe-chromosomal isolates) belong to a distinct cluster that is significantly distant from all the other cpb2 isolates (cpe-plasmid carrying and cpe-negative isolates). The isolates carrying a cpb2 appear to be rarely related, and are more variable than chromosomal cpe isolates. The results suggest that the cpe-positive outbreak isolates showed close genetic relation, whereas the cpb2-positive isolates revealed a wide genetic diversity.

A single case study using Contextual Support Model based music therapy intervention for a terminal pediatric patient (말기질환 아동을 위한 환경적 지지모델(The Contextual Support Model of Music Therapy)에 근거한 음악치료사례연구)

  • Jun, Mi Hye
    • Journal of Music and Human Behavior
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    • v.2 no.2
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    • pp.15-31
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    • 2005
  • Terminal pediatric patient include congenital metabolic abnormalities, chromosome aberrations, congenital anomalies, neuromuscular diseases and other incurable conditions as well as malignant tumors. One third of these diseases are cancers, and two thirds of cancers are lymphadenoma and leukemia. Terminal pediatric patient may feel fear, anger and frustration against treatment process in hospitals, lose control of themselves and stay in helplessness due to restrictions within controlled hospital environment. This study examined the relationship between hospitalized childrens behavior and music using Contextual Support Model of Music Therapya theory stating therapeutic music environments possess three elements of Structure, Autonomy support and Involvement, and increase childrens active engagement. Focused on these three therapeutic elements, this study analyzed music therapy cases of terminally ill children to examine the relationship between childrens behavior and music as environmental supportive medium in the environment. This study is on a single case case subject, a 10-year-old girl with acute lymphocytic leukemia. Nine sessions of music therapy activities were conducted and analyzed using qualitative method. Focusing on three therapeutic elements of Contextual Support Model, analysis of sessions was made on the basis of activities specifically designed for this study. Main music therapy activities included singing while playing musical instruments, singing while listening to music through CD player, making up songs, and searching for song lines. The findings of this study are as follows: a) in terms of Structure, music therapy suggested a direction for effective musical activities for terminally ill children by providing environments where child can sing, play musical instruments and make up songs; b) in terms of Autonomy support, music therapy encouraged childs voluntary participation by giving her chances to make choice while singing or playing musical instruments; c) in terms of Involvement, child could gain positive approval of the therapist and conduct musical activities in cooperation with the therapist. Music therapy serves as supportive medium in the environment, develops spontaneity and self-reliance in emotionally depressed children, and promotes voluntary attitudes in the restricted hospital environment. Based on the positive effects of music therapy, more studies need to be conducted with an approach to more various terminal pediatric patients.

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Application of a New Conjugation Method to Fish Pathogenic Bacteria Containing R Plasmid for the Analysis of Drug-Resistant Status in Aquaculture (새로운 conjugation 방법을 응용한 R plasmid 함유 어병세균의 분리와 양식장 내성균의 현황 분석)

  • Yoo Min Ho;Jeong Joon Beom;Kim Eun Heui;Lee Hyoung Ho;Jeong Hun Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.2
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    • pp.115-121
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    • 2002
  • To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.