• Title/Summary/Keyword: synthetic gene

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

  • 오봉국;손시환;이정구
    • Korean Journal of Poultry Science
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    • v.19 no.3
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    • pp.113-123
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    • 1992
  • This study was carried out to build up new synthetic egg Production lines which had sex linked gene for feather color sexing and had also superior combining ability for producing the best commercial chicks. In order to make autosexing layer line, the commercial layers which had Z$^{s}$ Z$^{s}$ and Z$^{s}$ W were mated. Among progeny, the chicks which had homozygote of silver gene and non-silver gene were selected for making dam and sire lines. Afterwards the closed flock breeding method was utilized to improve general performances of the each line. The performances of egg production in synthetic line were 161 day for age at sexual maturity, 219 eggs for total egg number to 60 weeks of age, 84% for hen-day egg production and 619 for average egg weight. There was no difference in egg production between new synthetic lines and imported breeds. In the analysis of genetic trends, the estimates of genetic parameter in the autosexing lines were similar to those of the general population of layer breeders. This results indicated the consistency of genetic variation from this selection.

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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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    • v.11 no.1
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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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    • v.39 no.6
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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
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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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
    • Proceedings of the Ginseng society Conference
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    • 1990.06a
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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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    • v.23 no.3
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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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    • v.24 no.12
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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.

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 (한국산 멧누에나방(Bombyx mandarina)에 있어서 난각유전자의 형질발현. II. 난각단백질의 합성과정)

  • 노시갑
    • Korean journal of applied entomology
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    • v.32 no.4
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    • pp.420-425
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    • 1993
  • The programme of silkrnoth chorion protem synthetIc changes is autonomous i. e.,it is imple I mented WIth normal kinetics by follicles cultured in isolation in a defined tIssue culture medium. On the basis of protem synthetic profiles, 8 stages of chorwgenesis are defined. Detail a analysis of the proteins of chorion of Bombyx mandarina, reveals more than 17 components by eletrophoretic mobility and synthetic kinetics. We have defined three abbreviated synthetlc s stages, based on the pattern of protein synthesized at early, middle, and late synthetic stages

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