• Title/Summary/Keyword: Gene combination

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Antimicrobial resistance and distribution of resistance gene in Enterobacteriaceae and Pseudomonas aeruginosa isolated from dogs and cats (개와 고양이에서 분리된 Enterobacteriaceae와 Pseudomonas aeruginosa의 항균제 내성 및 내성 유전자의 분포)

  • Cho, Jae-Keun;Kim, Jin-Hyun;Kim, Jeong-Mi;Park, Choi-Kyu;Kim, Ki-Seuk
    • Korean Journal of Veterinary Service
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    • v.36 no.3
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    • pp.171-180
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    • 2013
  • This study was carried out to investigate the antimicrobial resistance pattern and distribution of resistance gene in 44 Enterobacteriaceae and 21 Pseudomonas (P) aeruginosa isolated from hospitalized dogs and cats in animal hospital from 2010 to 2011 in Daegu. Among Enterobacteriaceae, Escherichia (E) coli was highly resistant to ampicillin (56.7%), followed by tetracycline (53.3%), cephalothin, streptomycine, sulfamethoxazole/trimethoprim, gentamicin and norfloxacin (40.0~43.3%). The remaining isolates of Enterobacteriaceae had high resistance to ampicillin (64.3%) and streptomycin (42.9%). Whereas, P. aeruginosa was low resistant to all antimicrobials tested (less than 15%). int I 1 gene was detected in 20 (57.1%) of 35 antimicrobial resistant Enterobacteriaceae and 2 (9.5%) of 21 P. aeruginosa., but int I 2 gene was not detected in all isolates. The eight resistance genes were found either alone or combination with other gene (s): $bla_{TEM}$, aadA, strA-strB, clmA, tetA, tetB, sul I and sul II. About 78% of integron-positive isolates were resistance to more than four antimicrobial agents. The findings suggest that class I integrons are widely distributed in E. coli among Enterobacteriaceae from dogs and cats and multi-drug resistance related to the presence of class I integrons. The prudent use of antimicrobials and continuous monitoring for companion animals are required.

Combination of Epstein-Barr Virus-Based Plasmid and Nonviral Polymeric Vectors for Enhanced and Prolonged Gene Expression

  • Choi, Hye;Park, Key Sun;Bae, Seon Joo;Song, Su Jeong;Kim, Kyoon Eon;Park, Jong-Sang;Choi, Joon Sig
    • Bulletin of the Korean Chemical Society
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    • v.33 no.11
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    • pp.3676-3680
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    • 2012
  • An Epstein-Barr virus (EBV)-based plasmid contains the EBV nuclear antigen 1 (EBNA1) gene and EBV replication origin (oriP) sequence. Since EBNA1 (the only EBV-encoded protein) is combined with oriP, it is replicated simultaneously with chromosomal DNA in human, primate, and canine cells and is faithfully segregated at a stable copy number upon cell division. Consequently, it can be used to stably express gene inserts over a prolonged time in target cells. We have previously shown that the polyamidoamine (PAMAM) dendrimer can be surface-modified with L-arginine. Arginine is present at a high frequency in the transactivator of transcription (Tat) sequences of human immunodeficiency virus (HIV). It presents high membrane permeability and permits effective transfer of DNA inside the cells. In this study, we constructed two kinds of recombinant DNA by inserting the luciferase gene and enhanced green fluorescence protein (eGFP) gene as reporter genes into the pCEP4 plasmid vector. We measured dynamic light scattering (DLS) and zeta potential after preparing PAMAM-based cationic polymer/EBV-based plasmid complexes. We performed transfection of HEK 293 cell lines with the polyplexes, and monitored luciferase activity and green fluorescence protein (GFP) expression. Our results show that PAMAM-based cationic polymer/EBV plasmid complexes provide enhanced and sustained gene expression.

Development of Gene-based Markers for the Allelic Selection of the Restorer-of-fertility Gene, Rfo, in Radish (Raphanus sativus)

  • Kim, Sunggil;Lim, Heerae;Cho, Kang-Hee;Park, Pue Hee;Park, Suhyung;Sung, Soon-Kee;Oh, Daegeun;Kim, Ki-Taek
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.194-204
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    • 2009
  • Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for $F_1$-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in $F_1$-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating $F_1$-hybrid cultivar development in radish.

Genetic Analysis of Quantitative Characters of Rice (Oryza sativa L.) by Diallel Cross (이면교배(二面交配)에 의한 수도량적(水稻量的) 형질(形質)의 유전분석(遺傳分析)에 관(關)한 연구(硏究))

  • Jo, Jae-seong
    • Korean Journal of Agricultural Science
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    • v.4 no.2
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    • pp.254-282
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    • 1977
  • To obtain information on the inheritance of the quantitative characters related with the vegetative and reproductive growth of rice, the $F_1$ seeds were obtained in 1974 from the all possible combinations of the diallel crosses among five leading rice varieties : Nongbaek, Tongil, Palgueng, Mangyeong and Gimmaze. The $F_1$'s including reciprocals and parents were grown under the standard cultivation method at Chungnam Provincial Office of Rural Development in 1975. The arrangement of experimental plots was randomized block design with 3 replications and 12 characters were used for the analysis. Analytical procedure for genetic components was followed the Griffing's and Hayman's methods and the results obtained are summarized as follows. 1. In all $F_1$'s of Tongil crosses, the longer duration to heading was due to dominant effect of Tongil and each $F_1$ showed high heterosis in delaying the heading time. It was assumed that non-allelic gene action besides dominant gene effect might be involed in days to heading character. However, in all $F_1$'s from the crosses among parents excluding Tongil the shorter duration was due to dominant gene action and the degree of dominance was partial, since dominance effects were not greater than the additive effect. The non-allelic gene interaction was not significant. Considering the results mentioned above, it was regarded that there were two kinds of Significantly different genetic systems in the days to heading. 2. The rate of heterosis was significantly different depending upon the parents used in the crosses. For instance, the $F_1$'s from Togil cross showed high rate of heterosis in longer culm. Compared to short culm, longer culm was due to recesive gene action and short culm was due to recesive gene action. The dominant gene effect was greater than the additive gene effect in culm length. The narrow sense of heretability was very low and the maternal effects as well as reciprocal effects were significantly recognized. 3. The lenght of the of the uppermost internode of each $F_1$ plant was a little lorger than these of respective parental means or same as those of parents having long internodes, indicating partial dominance in the direction of lengthening the uppermost internodes. The additive gene effects on the uppermost internode was greater than the dominance gene effect. The narrow as well as broad sense of heritabilities for the character of the uppermost internode were very high. There were significant maternal and reciprocal effect in the uppermost internode. 4. The gene action for the flag leaf angle was rather dominance in a way of getting narrower angle. However, in the Palgueng combinations, heterosis of $F_1$ was observed in both narrow and wide angles of the flag leaf. The dominant effects were greater than the additive effects on the flag leaf angle. There were observed also a great deal of non-allelic gene interacticn on the inheritance of the flag leaf angle. 5. Even though the dominant gene action on the length and width of flag leaf was effective in increasing the length or width of the flag leaf, there were found various degrees of hetercsis depending upon the cross combination. Over-dominant gene effect were observed in the inheritance of length of the flag leaf, while additive gene effects was found in the inheritance of the width of the flag leaf. High degree of heretabilities, either narrow or broad sense, were found in both length and width of the flag leaf. No maternal and reciprocal effect were found in both characters. 6. When Tongil was used as one parent in the cross, the length of panicle of $F_1$'s was remarkedly longer than that of parents. In other cross comination, the length of panicle of $F_1$'s was close to the parental mean values. Rather greater dominent gene effect than additive gene effect was observed in the inheritance of panicle length and the dominant gene was effective in increasing the panicle length. 7. The effect of dominant genes was effective in increasing the number of panicles. The degree of heterosis was largely dependent on the cross combination. The effect of dominant gene in the inheritance of panicle number was a little greater than that of additive genes, and the inheritance of panicle number was assumed to be due to complete dominant gene effects. Significantly high maternal and reciprocal effects were found in the character studied. 8. There were minus and plus values of heterosis in the kernel number per panicle depending upon the cross combination. The mean dominant effect was effective in increasing the kernel number per panicle, the degree of dominant effect varied with cross combination. The dominant gene effect and non-allelic gene interaction were found in the inheritance of the kernel number per panicle. 9. Genetic studies were impossible for the maturing ratio, because of environmental effects such as hazards delaying heads. The dominant gene effect was responsible for improving the maturing ratio in all the cross combinations excluding Tongil 10. The heavier 1000 grain weight was due to dominant gene effects. The additive gene effects were greater than the dominant gene effect in the 1000 grain weight, indicating that partial dominance was responsible for increasing the 1000 grain weight. The heritabilites, either narrow or broad sense of, were high for the grain weight and maternal or reciprocal effects were not recognized. 11. When Tongil was used as parent, the straw weight was showing high heterosis in the direction of increasing the weight. But in other crosses, the straw weight of $F_1$'s was lower than those of parental mean values. The direction of dominant gene effect was plus or minus depending upon the cross combinations. The degree of dominance was also depending on the cross combination, and apparently high nonallelic gene interaction was observed.

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Genetically Engineered Mouse Models for Drug Development and Preclinical Trials

  • Lee, Ho
    • Biomolecules & Therapeutics
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    • v.22 no.4
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    • pp.267-274
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    • 2014
  • Drug development and preclinical trials are challenging processes and more than 80% to 90% of drug candidates fail to gain approval from the United States Food and Drug Administration. Predictive and efficient tools are required to discover high quality targets and increase the probability of success in the process of new drug development. One such solution to the challenges faced in the development of new drugs and combination therapies is the use of low-cost and experimentally manageable in vivo animal models. Since the 1980's, scientists have been able to genetically modify the mouse genome by removing or replacing a specific gene, which has improved the identification and validation of target genes of interest. Now genetically engineered mouse models (GEMMs) are widely used and have proved to be a powerful tool in drug discovery processes. This review particularly covers recent fascinating technologies for drug discovery and preclinical trials, targeted transgenesis and RNAi mouse, including application and combination of inducible system. Improvements in technologies and the development of new GEMMs are expected to guide future applications of these models to drug discovery and preclinical trials.

Morphological and Molecular Phylogenetic Data of the Chinese Medicinal Fungus Cordyceps liangshanensis Reveal Its New Systematic Position in the Family Ophiocordycipitaceae

  • Wang, Yao;Dai, Yong-Dong;Yang, Zhong-Lin;Guo, Rui;Wang, Yuan-Bing;Yang, Zhu L.;Ding, Lei;Yu, Hong
    • Mycobiology
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    • v.49 no.4
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    • pp.297-307
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    • 2021
  • A cordycipitoid fungus infecting Hepialidae sp. in Nepal was supposed to be identical to Cordyceps liangshanensis, originally described from southwestern China, and thus, transferred to the genus Metacordyceps or Papiliomyces in previous studies. However, our multi-gene (nrSSU-nrLSU-tef-1α-rpb1-rpb2) phylogenetic and morphological studies based on the type specimen and additional collections of C. liangshanensis revealed that the fungus belongs to the genus Ophiocordyceps (Ophiocordycipitaceae). Therefore, a new combination O. liangshanensis was made, and a detailed description of this species was provided.

Effects of a Pueraria lobata-root based combination supplement containing Rehmannia glutinosa and aerobic exercise on improvement of metabolic dysfunctions in ovariectomized rats (갱년기 모델 유도 흰 쥐에서 갈근과 지황 복합물 및 유산소 운동이 대사이상 개선에 미치는 영향)

  • Oh, Sang A;Ok, Hyang Mok;Kim, Hye Jin;Lee, Won Jun;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.48 no.2
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    • pp.133-139
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    • 2015
  • Purpose: There is a fair amount of evidence indicating that increased risk of obesity and insulin resistance is associated with postmenopausal state, but can be modulated by diet and exercise. In this study, we explored whether a Pueraria lobata root-based supplement containing Rehmannia glutinosa (PR) and/or aerobic treadmill exercise can modify the metabolic changes associated with estrogen deficiency. Methods: Seventy rats were randomly assigned to the following groups for 8 weeks (n=10 per group): SHAM, sham-operated; PR0, ovariectomized (OVX) control; PR200, OVX with PR200 mg/kg B.W; PR400, OVX with PR400 mg/kg B.W; EPR0, OVX with exercise; EPR200, OVX with exercise and PR200 mg/kg B.W; EPR400, OVX with exercise and PR400 mg/kg B.W. Results: OVX induced significant increases in body weight, food intake, fat mass, LDL-cholesterol, and fasting blood glucose, confirming induction of menopausal symptoms. PR supplementation or exercise significantly suppressed the above mentioned changes through different regulatory elements in adipose tissue: PR supplement upregulated adiponectin gene expression and aerobic exercise upregulated adiponectin and insulin receptor gene expression and a combination of PR supplement and aerobic exercise showed an additive effect on adiponectin gene expression. Conclusion: Taken together, the results of this study suggest that PR supplement has a potential to provide health benefits in OVX rats through leptin and adiponectin secretion. In addition, the data suggest that combination of exercise and PR would have additive effects on metabolic dysfunction associated with estrogen deficiency.

Oxidation of fatty acid may be enhanced by a combination of pomegranate fruit phytochemicals and acetic acid in HepG2 cells

  • Kim, Ji Yeon;Ok, Elly;Kim, You Jin;Choi, Kyoung-Sook;Kwon, Oran
    • Nutrition Research and Practice
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    • v.7 no.3
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    • pp.153-159
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    • 2013
  • We investigated whether the combination of phytochemicals and acetic acid in the form of fruit vinegar provides an additive effect on changes of mRNA levels related to fatty acid oxidation in human hepatocyte (HepG2). Among the seven fruit vinegars (Rubuscoreanus, Opuntia, blueberry, cherry, red ginseng, mulberry, and pomegranate) studied, treatment of HepG2 with pomegranate vinegar (PV) at concentrations containing 1 mM acetic acid showed the highest in vitro potentiating effect on the mRNA expression levels of peroxisome proliferator-activated receptor ${\alpha}$, carnitinepalmitoyl transferase-1, and acyl-CoA oxidase compared to the control group (P < 0.05). Reversed-phase liquid chromatography in combination with quadrupole time-of-flight mass spectrometry analysis revealed four potential compounds (punicalagin B, ellagic acid, and two unidentified compounds) responsible for altered gene expression in HepG2 cells treated with PV as compared with the others. Further investigations are warranted to determine if drinking PV beverages may help to maintain a healthy body weight in overweight subjects.

Characterization of an Oxygen-Dependent Inducible Promoter Systems, the nar Promoter of Escherichia coli, and Gram negative host strains

  • Lee, Gil-Ho;Jo, Mu-Hwan;Lee, Jong-Won
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.762-766
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    • 2001
  • The nar promoter of Escherichia coli was known to induce maximally under anaerobic or microaerobic conditions in the presence of nitrate. In this study, the nar promoter was tested to see whether the expression level of a reporter gene which fused lacZ gene at nar promoter's downstream, in the some gram negative host strains(Agrobacterium, Pseudomonas and Rhizobium). A nar promoter system(Combination of nar promoter and gram negative strain) was grown under aerobic conditions to absorbance at 600 nm of nearly 2.0 and then, the nar promoter was induced by lowering DO to 1-2% with alternating microaerobic and aerobic condition in the fermentor cultures, using different gram negative hosts. For a wild type nar promoter (pNW61), it was possible to maintain production of ${\beta}-galactosidase$ activity per cell(specific ${\beta}-galactosidase$ activity) at 14,000, 9600, 45 Miller units in the presence of 1% nitrate. and for a nitrate - independent nar promoter (pNW618) at 12,000, 10,400 and 58 Miller units in the absence of nitrate ion, respectively.

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Schizosaccharomyces pombe rsml Genetically Interacts with spmex67, Which Is Involved in mRNA Export

  • Yoon, Jin-Ho
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.32-36
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    • 2004
  • We have previously isolated three synthetic lethal mutants from Schizosaccharomyces pombe in order to identify mutations in the genes that are functionally linked to spmex67 with respect to mRNA export. A novel rsm1 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMex1. The rsml gene contains no introns and encodes a 296 amino-add-long protein with the RING finger domain, a C3HC4 in the N-terminal half. The Δrsm1 null mutant is viable, but it showed a slight poly(A)$\^$+/ RNA accumulation in the nucleus. Also, the combination of Δrsm1 and Δspmex67 mutations confers synthetic lethality that is accompanied by the severe poly(A)$\^$+/ RNA export defect. These results suggest that rsm1 is involved in mRNA export from the nucleus.