• Title/Summary/Keyword: Resistant Genes

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Metabolic Engineering of the Brassinosteroid Biosynthetic Pathways (Brassinosteroid의 대사공학)

  • Lee, Mi-Ock;Song, Ki-Hong;Lee, Hyun-Kyung;Jung, Ji-Yoon;Choe, Vit-Nary;Choe, Sung-Hwa
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04b
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    • pp.69-75
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    • 2002
  • Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd/dwf3 were Shown to be blocked in $D^4$ reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bri1/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus it is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRI1 could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

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Metabolic Engineering of the Brassinosteroid Biosynthetic Pathways (Brassinosteroid의 대사공학)

  • Lee, Mi-Ock;Song, Ki-Hong;Lee, Hyun-Kyung;Jung, Ji-Yoon;Choe, Vit-Nary;Choe, Sung-Hwa
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.139-144
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    • 2002
  • Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd /dwf3 were shown to be blocked in D$^4$reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bril/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus it is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRIl could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

Characterization, detection and identification of transgenic chili pepper harboring coat protein gene that enhances resistance to cucumber mosaic virus

  • Seo, Sang-Gyu;Kim, Ji-Seong;Jeon, Seo-Bum;Shin, Mi-Rae;Kang, Seung-Won;Lee, Gung-Pyo;Hong, Jin-Sung;Harn, Chee-Hark;Ryu, Ki-Hyun;Park, Tae-Sung;Kim, Sun-Hyung
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.384-391
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    • 2009
  • Previously, two events (H15 and B20) of transgenic pepper (Capsicum annuum L.) that enhanced resistance to Cucumber mosaic virus (CMV) by the introduction of CMV coat protein (CP) gene were constructed. Presently, a single copy number of the CP gene was revealed in H15 and B20 by Southern blot. To predict possible unintended effects due to transgene insertion in an endogenous gene, we carried out sequencing of the 5'-flanking region of the CP gene and a Blastbased search. The results revealed that insertion of the transgene into genes encoding putative proteins may occur in the H15 and B20 transgenic event. Mutiplex polymerase chain reaction (PCR) for simultaneous detection and identification of transgenic pepper was conducted with a set of nine primers. Both transgenic event were differentiated from non-transgenic event by the presence of 267 bp and 430 bp PCR products indicative of CP gene specific primer pairs and primer pairs targeting the CP gene and 35S promoter. H15 and B20 uniquely possessed a 390 bp and 596 bp PCR product, respectively. The presence of a 1115 bp product corresponding to intrinsic pepper actin gene confirmed the use of pepper DNA as the PCR template. The primer set and PCR conditions used presently may allow the accurate and simple identification of CMV resistant transgenic pepper.

The Effect of the Transcriptional Regulation of Sp1 for TGF-β1 and CTGF Expression in Scar Formation (반흔형성 과정에서 Sp1 전사인자 조절에 의한 TGF-β1 및 CTGF의 발현)

  • Park, Dong Man;Sohn, Dae Gu;Han, Ki Hwan;Lee, Sun Young;Chae, Young Mi;Chang, Young Chae;Park, Kwan Kyu
    • Archives of Plastic Surgery
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    • v.33 no.1
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    • pp.39-45
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    • 2006
  • This study is to examine the relationship between TGF-b1 expression and CTGF expression, and to evaluate the effect of Sp1 blockade on the expression of TGF-b1, CTGF and extracellular genes, clones of fibroblasts stably transfected with Sp1 decoy ODN. R-Sp1 decoy ODN was highly resistant to degradation by nucleases or serum, compared to the linear or phosphorothioated-Sp1 decoy ODN. Skin wounds were created on the back of 36 anesthetized rats. They were divided into four groups-the rats with normal skin, with wounded skin without decoy, with wounded skin injected with R-Sp1 decoy, and with wounded skin injected with mismatched R-Sp1 decoy, respectively. Skins were collected at 3rd, 5th, 7th, 14th day after wounding. Cellular RNA was extracted by RT-PCR analysis. TGF-${\beta}1$ and CTGF were deeply related with skin fibrosis during scar formation and it appeared that TGF-${\beta}1$ may cause the induction of CTGF expression. R-Sp1 decoy ODN inhibited TGF-${\beta}1$ and CTGF expression both in cultured fibroblasts and in the skin of rats. These results indicate that targeting Sp1 with R-type decoy efficiently blocks extracellular matrix gene expression, and suggest an important new therapeutic approach to control the scarring in normal wound healing and fibrotic disorders.

Characterization of Plasmid-Mediated SHV-11 β-lactamase Gene of Klebsiella pneumoniae Isolated from the Clinical Specimens (임상검체로부터 분리한 플라스미드 매개성 SHV-11 β-lactamase 유전자의 특성)

  • Kim, Yun-Tae;Lee, Sang-Hoo;Jang, Ji-Hyun;Kim, Tae-Un;Choi, Seok-Cheol;Baik, Hyung-Suk;Lee, Kyoung-Ryul;Yoon, Hye-Ryoung;Kim, Young-Jin
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1718-1723
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    • 2009
  • In this study, we characterized extended-spectrum $\beta$-lactamase (ESBL)-producing Enterobacteriaceae isolated from clinical specimens in Korea and found two strains harboring plasmid-mediated $bla_{SHV-11}$, Klebsiella pneumoniae. First, the isolates were detected using the Vitek system and confirmed by the double-disk synergy test. The classification of gene coding for ESBL was also performed by polymerase chain reactions and followed by DNA sequencing. The transmission of genes was confirmed by transconjugation and transformation. Resistant expression of transformants was determined by broth microdilution minimal inhibitory concentration test. Genotypic analysis revealed that one strain harbored the $bla_{TEM-1}$, $bla_{SHV-11}$ and $bla_{CTX-M-15}$ and the other strain harbored the $bla_{SHV-11}$ and $bla_{CTX-M-15}$. They showed high resistance to oxyiminocephalosphorins (3rd-generation cephalosporins), while the transformant containing only $bla_{SHV-11}$ did not show any resistance to the antibiotics.

A Three-year Survey on Korean Consumer's Awareness, Perception and Attitudes toward Genetically-modified Foods; Years 2000-2002 (유전자재조합식품의 인지도 및 수용도에 대한 연차별 비교)

  • Kim, Myung-Hee;Kim, Jay-Wook;Chae, Kyung-Yeon;Park, Se-Won;Kim, Youn-Soon;Kyung, Kyu-Hang
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1155-1161
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    • 2003
  • A three-year (2000-2002) survey on consumer's awareness and perception of genetically-modified (GM) foods was conducted on random samples of Korean consumers. More than 65% of the respondents were exposed to some information related to GM foods. The greatest benefit of the development of GM foods was thought to be their remedy of potential food shortages in the future. More than 90% of Korean consumers wanted GM foods to be labeled. About 18% of the respondents would buy GM foods voluntarily, whereas over 46% would not until they knew more about the product. Only 39% of Korean consumers were found to have realized that food items origination from plants contained genes. More consumers responded that they would not buy herbicide-resistant GM soybean and buy vitamin-enriched GM soybean instead. Many Korean consumers appeared to make decisions of acceptance or rejection of GM foods not on the basis of biotechnology, but on the basis of the word(s) used to describe the products, such as herbicide and vitamin. Only 4% of Korean consumers responded that GM foods were the greatest threat to the safety of Korean foods.

Effect of plasmid curing on the 2, 3-dihydroxybenzoic acid production and antibiotic resistance of Acinetobacter sp. B-W (Acinetobacter sp. B-W의 2, 3-dihydroxybenzoic acid 생산과 항생제 저항성에 미치는 플라스미드 제거 효과)

  • Kim, Kyoung-Ja;Kim, Jin-Woo;Yang, Yong-Joon
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.254-259
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    • 2016
  • Acinetobacter sp. B-W producing siderophore, 2, 3-dihydroxybenzoic acid (DHB) was analyzed for plasmid content. Strain B-W harbored plasmid of 20 kb in size. Growth at $43^{\circ}C$ was effective in producing mutant cured of plasmid of strain B-W. This mutant lost the ability to produce 2, 3-DHB. Formation of siderophore halos on the chrome azurol S (CAS) agar medium was not detected by cured strain B-W. pHs of supernatants of wild type strain B-W and cured mutant grown in glucose and $MnSO_4$ containing medium at $28^{\circ}C$ for 3 days were 4.5 and 8.5, respectively. Antibiotic resistance against ampicillin, actinomycin D, bacitracin, lincomycin, and vancomycin was lost in cured mutant. Plasmid curing of strain B-W resulted in drastic reduction of minimal inhibitory concentration (MIC) of several antibiotics. E. coli $DH5{\alpha}$ was transformed with plasmid isolated from strain B-W. The transformant E. coli $DH5{\alpha}$ harbored a plasmid of the same molecular size as that of the donor plasmid. Transformant E. coli $DH5{\alpha}$ produced 2, 3-DHB and contained antibiotic resistant ability. Thus a single plasmid of 20 kb seemed to be involved in 2, 3-DHB production. Genes encoding resistance to antibiotics were also supposed to be located on this plasmid.

Characteristics of Haploid Plants Derived from Interspecific Cross between Cytoplasmic Male-sterile Tobacco $F_1$(Nicotiana tabacum) and Nicotiana africana (연초(Nicotiana tabacum L.) 새포질 웅성불임 $F_1$과 Nicotiana africana의 종간 교배에 의한 반수체 식물의 특성)

  • 정윤화;금완수;조명조
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.5
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    • pp.649-654
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    • 1995
  • This study was conducted to investigate the utility and agronomic characteristics and use of cytoplasmic. male-sterile (CMS) haploid plants derived from interspecific cross between (male-sterile NC82$\times$burley21) F$_1$ plant (Nicotiana tabacum L.) and Nicotiana africana. Abundant seeds of high germinability were obtained when Nicotiana tabacum (cytoplasmic male-sterile F$_1$ plants) is pollinated by Nicotiana africana. Most of seedlings died at the cotyledonary stage. The remaining seedlings are viable F$_1$ hybrids or maternal haploids that can be easily distinguished. Number of interspecific Fl hybrids and matermal haploids per capsule obtained from the interspecific cross between cytoplasmic male-sterile tobacco F$_1$ plants and N.africana yielded 2.2 and 0.5 plants, respectively. Out of 149 CMS haploid plants obtained from interspecific cross, 102 plants showed green type while the others were yellow type for leaf and stem. This results agreed with the genetic ratios expected among haploid plants from the F$_1$ hybrids heterozygous for two recessive genes of yellow color of burley tobacco plant. Out of 83 CMS haploid plants inoculated with TMV, 48 plants showed resistant, while the others was susceptible. It agreed with the expected genetic ratios for a single dominant gene of TMV resistance. CMS haploid plant will be useful as a source material for breeding of CMS doubled haploid lines

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QTLs Identification and Confiirmation of Field Resistance to Leaf Blast in Temperate japonica Rice (Oryza sativa L.)

  • Cho, Young-Chan;Kwon, Soon-Wook;Suh, Jung-Pil;Kim, Jeong-Ju;Lee, Jeom-Ho;Roh, Jae-Hwan;Oh, Myung-Kyu;Kim, Myeong-Ki;Ahn, Sang-Nag;Koh, Hee-Jong;Yang, Sae-Jun;Kim, Yeon-Gyu
    • Journal of Crop Science and Biotechnology
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    • v.11 no.4
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    • pp.269-276
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    • 2008
  • Field resistance is defined as the resistance that allows effective control of a parasite under natural field condition and is durable when exposed to new races of that parasite. To identify the genes for field resistance to rice blast, quantitative trait loci (QTLs) conferring the resistance for races and blast nursery screening in japonica rice cultivars were detected and mapped using SSR markers. QTL analysis was carried out in 190 RILs population from the cross between Suweon365 (moderately resistant) and Chucheong (highly susceptible). Twelve QTLs against nine blast races inoculated were detected on chromosomes 1, 2, 4, 6, 7, 11 and 12. They explained from 5.1% to 34.9% of total phenotypic variation. Eight QTLs against blast nursery screening in four regions for three years were detected on chromosomes 1, 2, 4, 11 and 12. The phenotypic variation explained by each QTL ranged from 4.3% to 37.7%. Three chromosome segment substitution lines (CSSLs) of $BC_2F_6$ by backcross method were developed to transfer the QTLs into the susceptible cultivar Chucheong as a recurrent parent. A CSSL4-1 containing two QTLs qLB6.2 and qLB7 against blast races showed to the reaction of 6 to 7 at blast nursery in two regions for two years. The CSSL4-2 and CSSL93 containing QTLs, qLB11.2 and qLB12.1 of the resistance against leaf blast in blast nursery screening, respectively, had enhanced the resistance for blast nursery screening across two regions and in two years.

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Inhibition of Oncogenes Affects the Expression of NKG2D Ligands in Cancer Cells (k-ras와 c-myc, wnt 억제에 의한 NKG2D 리간드의 발현변화)

  • Heo, Woong;Lee, Young Shin;Bae, Jaeho
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1216-1222
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    • 2013
  • NK cells are lymphoid immune cells that participate in innate immunity to protect against foreign pathogens and transforming cells. It is known that the activity of NK cells is regulated by a balance between activating and inhibitory signals rather than specific antigens. One important activating signal is mediated by the NKG2D receptor, which recognizes NKG2D ligands on cancer cells. Therefore, tumor cells that express sufficient amounts of NKG2D ligands could be eliminated by NKD2D+ cells, including NK cells. Oncogenes drive tumor cells to apoptosis resistant and uncontrolled proliferation by altered expression of many critical genes. Therefore, the expression of NKG2D ligands may be affected by oncogenes. This study focused on increasing the susceptibility of cancer cells to NK cells by regulating the expression of NKG2D ligands influenced by three oncogenes: k-ras, wnt, and c-myc. We demonstrated that inhibition of k-ras and c-myc increased the expression of NKG2D ligands and enhanced the susceptibility of cancer cells to NK cells. On the contrary, inhibition of the wnt pathway decreased MICA and ULBP1 transcripts. Although the decreased transcription of NKG2D ligands by inhibition of the wnt pathway, surface proteins of NKG2D ligands were not changed, and the susceptibility of HCT-116 cells was unaffected. The results demonstrate that the transcription of NKG2D ligands are regulated deferentially by the k-ras, c-myc, and wnt pathways and that the cytotoxicity of NK cells solely depends on the amount of surface NKG2D ligands.