• 제목/요약/키워드: synthetic gene

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성 감별 유전자를 도입한 다산계계통 신품종 육종에 관한 연구 I. 반생유전계통 조성 (Breeding of New Synthetic Egg Production Line in Domestic Chicken by Intlroducing Sex Linked Gene. I. Production of the Autosexing Breed)

  • 오봉국;손시환;이정구
    • 한국가금학회지
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    • 제19권3호
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    • pp.113-123
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    • 1992
  • 본 연구는 반성은색유전자(S:silver gene)를 도입하여 성 감별용 산란종계 계통으로 육성하여 이를 모계통으로 이용함으로서 깃털에 의한 자웅감별이 가능할 뿐만 아니라 잡종강세를 이용한 우수실용계를 작출하고자 수행되었다. 암수 감별용 계통 조성을 위하여서는 현재 시판되고 있는 백색 실용계 수컷(Z$^{s}$ Z$^{s}$ ) 및 갈색 실용계 암컷(Z$^{s}$ W)을 이용하여 이들 중 우수개체를 선발하여 교잡시킴으로서 생산된 개체중 유전적 조성이 은색유전자를 동형(homo)으로 가진 개체들을 모계통으로 육성하고, 부계통 조성을 위하여서는 갈색유전자를 동형으로 가진 개체를 선발 육성하므로서 감별의 원종계를 작출하였다. 이들 조성된 계통을 이용하여 생산능력이 우수한 계통으로 육종하기 위하여 폐쇄군 육종방법(clossed flock breeding)을 사용하였다. 합성종 계계의 산란능력 검정 결과 60주령까지의 산란기록에 근거한 주요 경제능력은 초산 일령 161일, 60주령시 산난수 219개, 산난율 84% 평균난중 61g으로 외국에서 수입되는 우수종계에 버금가는 성적을 나타내었다. 또한 깃털에 의한 자웅감별이 가능할 뿐만 아니라 산난능력도 우수한 실용성 있는 암수 감별용 합성종을 육성 할 것으로 판단된다. 합성종 계통의 유전분석 결과 이들의 유전모수가 정상집단의 유전모수와 유사한 추정치를 보임으로서 특정계통 조성을 위한 선발에 기인된 유전변이의 변화는 거의 없음을 시사한다.

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Biodistribution and Hemolysis Study of Terplex Gene Delivery System in Mice

  • Oh, Eun-Jung;Shim, Jin-young;Kim, Jin-Seok
    • Macromolecular Research
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    • 제11권1호
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    • pp.19-24
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    • 2003
  • Polymeric gene delivery system attracts profound attention as it shows less toxicity, versatility, and reasonable gene expression efficiency. Terplex system, a synthetic biopolymeric gene delivery system consisting of stearyl poly-L-lysine (stearyl-PLL) and low density lipoprotein (LDL) was evaluated for its body distribution of gene expression of exogenously administered pDNA after tail-vein injection in mice. Kidney and spleen are two major organs with highest gene expression, whereas liver and heart showed marginal gene expression among the organs examined. Hemolytic effect of the terplex system was evaluated using human red blood cells, where terplex system did not cause significant hemolysis at the concentrations above the experimental ranges, although unmodified PLL or stearyl-PLL without LDL did. Serum stability of terplex system against enzymatic degradation was also significantly enhanced, presumably due to the steric stabilization from the polymers. Based on these findings and along with its high in vitro transfection efficiency, terplex system could serve as a safe and efficient polymeric gene delivery system with many applications for the in vivo gene therapy.

Overexpression of Heat Shock Factor Gene HsfA3 Increases Galactinol Levels and Oxidative Stress Tolerance in Arabidopsis

  • Song, Chieun;Chung, Woo Sik;Lim, Chae Oh
    • Molecules and Cells
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    • 제39권6호
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    • pp.477-483
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    • 2016
  • Heat shock factors (Hsfs) are central regulators of abiotic stress responses, especially heat stress responses, in plants. In the current study, we characterized the activity of the Hsf gene HsfA3 in Arabidopsis under oxidative stress conditions. HsfA3 transcription in seedlings was induced by reactive oxygen species (ROS), exogenous hydrogen peroxide ($H_2O_2$), and an endogenous $H_2O_2$ propagator, 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB). HsfA3-overexpressing transgenic plants exhibited increased oxidative stress tolerance compared to untransformed wild-type plants (WT), as revealed by changes in fresh weight, chlorophyll fluorescence, and ion leakage under light conditions. The expression of several genes encoding galactinol synthase (GolS), a key enzyme in the biosynthesis of raffinose family oligosaccharides (RFOs), which function as antioxidants in plant cells, was induced in HsfA3 overexpressors. In addition, galactinol levels were higher in HsfA3 overexpressors than in WT under unstressed conditions. In transient transactivation assays using Arabidopsis leaf protoplasts, HsfA3 activated the transcription of a reporter gene driven by the GolS1 or GolS2 promoter. Electrophoretic mobility shift assays showed that GolS1 and GolS2 are directly regulated by HsfA3. Taken together, these findings provide evidence that GolS1 and GolS2 are directly regulated by HsfA3 and that GolS enzymes play an important role in improving oxidative stress tolerance by increasing galactinol biosynthesis in Arabidopsis.

Production of the taste-modifying protein, miraculin, in transgenic lettuce

  • Ezura, Hiroshi;Sun, Heyon-Jin
    • 한국식물생명공학회:학술대회논문집
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    • 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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    • pp.126-131
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    • 2005
  • Richadella dulcifica, a native shrub in tropical West Africa, gives red berries that have the unusual property of modifying a sour taste into a sweet taste. The red berries contain a taste-modifying protein named miraculin. A synthetic gene encoding miraculin was placed under the control of constitutive promoters and transferred to lettuce. High expression of miraculin was obtained, with accumulation of up to 1% total soluble protein in lettuce leaf. In addition, the miraculin expressed in lettuce possesses a taste-modifying activity.

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Inductive Effects of Ginseng Saponins on the Rat LDH A-gene and the Synthetic rate of Hepatocyte DNA in Regenerating Rat Liver Cells

  • Yoo, Kye-Jin;Lee, Kwang-Youl;Lee, Seung-Ki
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1990년도 Proceedings of International Symposium on Korean Ginseng, 1990, Seoul, Korea
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    • pp.58-64
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    • 1990
  • The effects of ginseng saponins, G-Rbl and G-Rc on the rat liver LDH A-gene transcnptional activity was investigated during pro-replicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl of G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 ma/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5·hours after partial hepatectomy. Dose dependent effect of G-Rbl and G-Rc (1-25 mg/100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal in- creases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc However, when the administration doses of G-Kbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rb 1 and G·Rc have stimulatory effect at the lower concentration (1 mg/100g B.W) and inhibitory effect at the higher concentration (20 moi loos 5.W) on the LDH A-gene transcription during regeneration of rat liver, Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA synthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G·Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 ug and 100 ug/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen enhancer activity for the hepatocyte proliferation during rat liver regeneration period. Keywords Inductive effects of ginsenosides, G-Rb, -Rc, and -Re, rat LDH A-gene transcription, the sin thetic rate of proteins and DNA in regeneration rat liver.

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Development of a Genome-Wide Random Mutagenesis System Using Proofreading-Deficient DNA Polymerase ${\delta}$ in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Oh Cheol;Kim, Sang-Yoon;Hwang, Dong Hyeon;Oh, Doo-Byoung;Kang, Hyun Ah;Kwon, Ohsuk
    • Journal of Microbiology and Biotechnology
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    • 제23권3호
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    • pp.304-312
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    • 2013
  • The thermotolerant methylotrophic yeast Hansenula polymorpha is attracting interest as a potential strain for the production of recombinant proteins and biofuels. However, only limited numbers of genome engineering tools are currently available for H. polymorpha. In the present study, we identified the HpPOL3 gene encoding the catalytic subunit of DNA polymerase ${\delta}$ of H. polymorpha and mutated the sequence encoding conserved amino acid residues that are important for its proofreading 3'${\rightarrow}$5' exonuclease activity. The resulting $HpPOL3^*$ gene encoding the error-prone proofreading-deficient DNA polymerase ${\delta}$ was cloned under a methanol oxidase promoter to construct the mutator plasmid pHIF8, which also contains additional elements for site-specific chromosomal integration, selection, and excision. In a H. polymorpha mutator strain chromosomally integrated with pHIF8, a $URA3^-$ mutant resistant to 5-fluoroorotic acid was generated at a 50-fold higher frequency than in the wild-type strain, due to the dominant negative expression of $HpPOL3^*$. Moreover, after obtaining the desired mutant, the mutator allele was readily removed from the chromosome by homologous recombination to avoid the uncontrolled accumulation of additional mutations. Our mutator system, which depends on the accumulation of random mutations that are incorporated during DNA replication, will be useful to generate strains with mutant phenotypes, especially those related to unknown or multiple genes on the chromosome.

Characterization of a Lichenase Isolated from Soil Metagenome

  • Kim, Sang-Yoon;Oh, Doo-Byoung;Kwon, Ohsuk
    • Journal of Microbiology and Biotechnology
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    • 제24권12호
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    • pp.1699-1706
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    • 2014
  • A lichenase gene (mt-lic) was identified for the first time through function-based screening of a soil metagenomic library. Its deduced amino acid sequence exhibited a high degree of homology with endo-${\beta}$-1,3-1,4-glucanase (having both lichenase and chitosanase activities), encoded by the bgc gene of Bacillus circulans WL-12. The recombinant lichenase overexpressed and purified from Escherichia coli was able to efficiently hydrolyze both barley ${\beta}$-glucan and lichenan. The enzyme showed maximal activity at a pH of 6.0 at $50^{\circ}C$, with Azo-barley-glucan as the substrate. The metal ions $Mn^{2+}$, $Mg^{2+}$, $Ca^{2+}$, and $Fe^{2+}$ enhanced the enzymatic activity, whereas the $Cu^{2+}$ and $Zn^{2+}$ ions inhibited the enzymatic activity. The $K_m$ and $V_{max}$ values of the purified lichenase were determined to be 0.45 mg/ml and 24.83 U/min/mg of protein, respectively.

한국산 멧누에나방(Bombyx mandarina)에 있어서 난각유전자의 형질발현. II. 난각단백질의 합성과정 (Chorion Gene Expression in the Cellular Differentiation and Accumulation of Chorion Protein Synthesis of Silkmoth, Bombyx mandarina(Lepidoptera: Bombycidae). II. Synthesis Programme of Chorion Proteins)

  • 노시갑
    • 한국응용곤충학회지
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    • 제32권4호
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    • pp.420-425
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    • 1993
  • 멧누에나방(Bombyx mandarina) 난각유전자의 형질발현기구를 규명하기 위한 연구의 일환으로 각형성과정에 있어서 난각단백질의 합성과정을 분석하였다. 난각단백질의 합성과정을 여포세포의 기난관배양에 의한 in vitro합성계에 의하여 분석한 결과, 합성과정은 8단계로 나눌 수 있었으며 SDS-PAGE 분석에 의해 17개 이상의 단백질 band로 분리되었다. 멧누에 난각단백질은 합성시기에 따라 3단계로 구분되며 초기에는 비교적 고분자 성분이, 합성의 중후기에는 비교적 저분자성분이 합성되는 등 난각단백질의 합성은 시기특이적으로 조절되었다.

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