• Title/Summary/Keyword: Polyethylene Glycol

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Regeneration of Fertile Transgenic Rice Plane from a Korean Cultivar, Nakdongbyeo (한국 재배종 낙동벼에서 임성 형질전환식물체의 재분화)

  • Soo In LEE;Hyun Jin CHUN;Chae Oh LIM;Jeong Dong BAHK;Moo Je CHO
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.175-182
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    • 1995
  • Rice is one of the most successful monocot in regenerating fertile and genetically stable transgenic plants. However there is no report of a rice line developed in Korea that can be used for regeneration of fertile and genetically stable transformants. In this paper we first demonstrate that a Korean variety Nakdongbyeo, is suitable to obtain transgenic rice plants. Protoplasts from embryogenic suspension cultures were co-transformed with HPT (hygromycin phosphotransferase) and GUS ($\beta$-glucuronidase) genes in separate plasmids in the presence of PEG (polyethylene glycol). In 5 independent experiment, the average frequency of calli showing hygromycin resistance were 1.73%. Plantlets were regenerated from the Hy $g^{R}$ calli. The average efficiency of plantlet regeneration was apprbximately 27%. Based on the GUS activities of hygromycin resistant calli, ca.35% of the resistant calli carried active GUS genes. The R0 transgenic plantlets were grown to maturity and Rl seeds were obtained. By examining the in siぉ activity of GUS in Rl seeds and seedlings, we confirmed that the GUS transgene driven by a CaMV 35S (cauliflower mosaic virus) promoter showed proper expression patterns. We also confirmed Mendelian segregation of the HPT transgene in the Rl generation.n.

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Effect of Osmotic and Solid Matrix Priming to Improve Germination and Early Growth of Carrot and Onion Seed (Osmotic Priming 및 Solid Matrix Priming 처리에 의한 당근과 양파종자의 발아증진과 초기생육에 미치는 영향)

  • 강점순
    • Journal of Bio-Environment Control
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    • v.13 no.3
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    • pp.135-142
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    • 2004
  • The objective of this study was carried out to determine which method is effective for improving seed germination in carrot and onion among osmotic or solid matrix priming. Seeds were osmotic primed with -0.5 MPa polyethylene glycol 8000 (PEG) solution and solid matrix primed with Micro Cel E (seed, Micro Cel E, and water mixed with the ratio of 4.0:2.0:8.0 by weight for carrot, and the ratio of 4.0:1.2:5.0 for onion at $20^{\circ}C$ for 4 days). Seeds were rapidly absorbed water after osmotic priming and solid matrix priming treatment. The moisture content of solid matrix primed seeds was lower than that of osmotic primed seeds in carrot and onion. Osmotic priming and solid matrix priming did not influenced on increasing of percent germination, but $T_{50}$ of treated seeds was shorter than those of untreated seeds at four temperature regimes. The effects of osmotic or solid matrix priming in reducing the $T_{50}$ was greater when the seeds were germinated at $15^{\circ}C$ than at $25^{\circ}C$ and $30^{\circ}C$ temperature. Solid matrix primed seeds germinated faster than osmotic primed seeds at all temperature in both carrot and onion. After priming, surface-dried seeds germinated faster than dried-back seeds in carrot and onion. Emergence speed of carrot and onion seeds was faster than untreated seeds by osmotic priming or SMP treatment although the final emergence percentage was not significantly influenced. On the other hand, early growth was not significantly influenced by osmotic priming or SMP treatment of carrot and onion seeds.

Interspecific Hybridization between Pleurotus ostreatus and Pleurotus sajor-caju by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리와 여름느타리버섯의 종간(種間) 교잡(交雜))

  • Yoo, Young-Bok;Lee, Haing-Sook
    • The Korean Journal of Mycology
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    • v.22 no.4
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    • pp.378-385
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    • 1994
  • Interspecific somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus and Pleurotus sajor-caju. The fusion products between incompatible strains did not form clamp connections. Fruiting body of the clampless fusants was induced by light-dark cycle on saw-dust-rice bran substrate in glass bottles. Out of them, seven somatic hybrids produced fruiting bodies of intermediate morphology of the two species. Light and low temperature were the initiating factors for the development of clamped hyphae from the clampless mycelial colonies. All of these basidiocarps had clamp connections. Eight fusants from the six crosses were analysed with the segregation of genetic characters by random spore isolates. In the three combinations, unexpected alleles were shown. Somatic hybrid between P188 (P. ostreatus 2-1 + P. sajor-caju 2-53) and P. florida 2-3 by triple cross produced fruiting bodies similar to those of fusant between P. ostreatus and P. florida. All the genetic charaters from the three strains were shown to segregate and recombine.

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A Study on the Dimensional Stability of Archaeological Waterlogged Salix koreensis Andersson Treated with Recycled PEG (재활용 PEG를 이용한 수침 고버드나무의 치수안정화 연구)

  • Yang, Seok-Jin;Lee, Soo;Kim, Jong-Hwa
    • Journal of the Korean Applied Science and Technology
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    • v.31 no.4
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    • pp.635-641
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    • 2014
  • Archaeological waterlogged woods require a process of dimensional stabilization for their conservation. PEG is the most widely used in the conservation of archaeological waterlogged wood. One of the easiest and commonly used methods is the impregnation of 40% polyethylene glycol followed by vacuum freeze drying. However, the waste fluid produced from the PEG treatment is black in color and has a severe odor due to the organic matter extracted from the wood. Thus It cannot be recycled and it was just thrown out. Color of waste fluid can be decolored with oxidation reaction by hydrogen peroxide. Properties of PEG before and after preservation treatment, and after oxidation with $H_2O_2$ were not changed. Dimensional stability of archaeological waterlogged Salix koreensis Andersson was studied with pure or recycled PEG. The ratio of impregnation solutions were 10:0, 7:3, 5:5, 3:7, 0:10 (pure PEG : recycled PEG). Impregnation process was carried out by putting the wood specimens 10% PEG solution for 5days, 20% for 5 days, 30% for 5 days finally 40% for 5 days. All of the specimens showed the weight change rate of 25%. SEM results provided that the dimensional change of were less than 4% PEG impregnated specimens. Comparing with pure PEG impregnation system, conservation precess mixed PEG also showed no significant changes. Conclusively, the recycled PEG can be used for archeological waterlogged wood conservation precess.

Process Optimization for Concentration and Stabilization of Recombinant Endoxylanase Expressed in Bacillus subtilis (Bacillus subtilis에서 발현된 재조합 Endoxylanase 농축과 안정화 공정의 최적화)

  • Choe, Yeong-Rok;Park, Jeong-Ha;;Kim, Yeong-Man;Gwon, Hyeon-Ju;Kim, Byeong-U
    • Microbiology and Biotechnology Letters
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    • v.32 no.4
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    • pp.322-327
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    • 2004
  • A strong constitutive PJH promoter from Bacillus sp. was applied to overexpress the endoxylanase gene (639 bp) in Bacillus subtilis. The expression plasmid, pJHKJ4, was designed to contain the $P_{JH}$ promoter and open reading frame of endoxylanase including its own promoter. The plasmid was introduced into B. subtilis DB431 and the resulting transformant was grown on LB glucose medium. At the end of cultivation, the endoxylanase activity in the culture supernatant reached about 140 DIm!. The enzyme in the supernatant was concentrated by ultrafiltration (MW cut-off 10 kDa and 30 kDa) and ammonium sulfate precipitation. For the concentration of the enzyme, ultrafiltration was more efficient than 70% ammonium sulfate precipitation. The stabilization of concentrated enzyme solution at $50^{\circ}C$ was examined with various stabilizers such as NaCI, glycerol, polyethylene glycol, sorbitol, and $CaCI_2$. The most effective stabilizers were found to be NaCI and $CaCI_2$.

L-Methionine Production by Protoplast Fusion of Brevibacterium flavum ATCC 14067 and Corynebacterium glutamicum ATCC 13032 (Brevibacterium flavum ATCC 14067과 Corynebacterium glutamicum ATCC 13032의 원형질체 융합에 의한 L-Methionine의 생산)

  • Bin, Jae-Hoon;Chung, Soo-Ja;Shin, Dong-Bun;Ryu, Beung-Ho
    • Korean Journal of Food Science and Technology
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    • v.23 no.5
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    • pp.561-567
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    • 1991
  • This study was designed to investigate the productivity of L-methionine by the method of protoplast fusion between Brevibacterium flavum ATCC 14067 and Corynebacterium glutamicm ATCC 13032, and then L-methionine production was performed to continuous fermentation using the immobilized fusant cells. Mutants B. flavum K 104($thr\;met\;Km^{r}\;Et^{r}\;Sm^{r}\;Tm^{r}\;as\;genetic\;marker$) and C. glutamicum B 70($thr\;Hos\;Km^{r}\;Et^{r}\;Sm^{r}\;Tm^{r}as\;genetic\;marker$) were isolated by MNNG treatment. On the other hand, protoplast of mutants were formed to treat with lysis solution containing $500{\mu}g/ml$ of lysozyme. The ratios of protoplast formation and regeneration were 99% and $64{\sim}66%$ respectively. Fusion frequency between B. flavum K 104 and C. glutamicum B 70 showed the $3.5{\times}10^{5}$ in the 35% polyethylene glycol(PEG6000) containing 3% PVP solution. The productivity of L-methionine by fusant BFCG 37 immobilized with sodium alginate was 0.89 g/l the batch fermentation and was $18.75mg/^{1}hr\;^{1}$ on the continuous fermentation at $30^{\circ}C$ for 72 hr.

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Effect of kinds and concentrations of osmoticum on somatic embryo induction and germination from suspended embryogenic cell in Larix kaempferi (낙엽송(Larix kaempferi) 현탁배양된 배발생세포로부터 체세포배 유도 및 발아를 위한 삼투압제 종류 및 농도 효과)

  • Kim, Yong-Wook
    • Journal of Plant Biotechnology
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    • v.40 no.3
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    • pp.141-146
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    • 2013
  • This study was conducted to examine suspended embryogenic cells growth with days of culture, effects of various kinds/concentrations of osmoticum for induction of somatic embryos (SEs), following somatic embryos germination or plantlet regeneration. The proliferation pattern of embryogenic cells in suspension culture is characterized by settled cells volume (SCV) increased with the duration of culture with marked the maxium of SCV (10.1 ml) in 18 days of culture, however the SCV of cells gradually decreased after that. In comparison of kinds/concentrations of osmoticum on somatic embryo induction, the highest induction number (352.3/g FW) of the SE was showed in 0.2 M sucrose, in addition, we also observed some effects with treatments of 0.2 M maltose (203.7) and 0.3 M maltose (193.7), respectively. However, no somatic embryos produced in treatments of 7.5% PEG plus 0.15 M sucrose or maltose. In comparison of germination efficiency of SEs which occurred from the treatments of various kinds/ concentrations of osmoticum, the highest induction frequency of cotyledon (25.2%) was obtained from SEs that produced 0.3 M maltose, however, the best occurrence rates of hypocotyl (39%), radicle (30.3%) and plantlet regeneration (3.5%) were observed from the 0.2 M sucrose treatment, respectively.

Interspecific protoplast Fusion and Sexuality in Pleurotus (느타리의 종간(種間) 원형질체(原形質體) 융합(融合) 및 유성(有性))

  • Yoo, Young-Bok;You, Chang-Hyun;Park, Yong-Hwan;Lee, Yeon-Hee;Chang, Kwon-Yawl;Peberdy, John F.
    • The Korean Journal of Mycology
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    • v.15 no.3
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    • pp.135-141
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    • 1987
  • Interspecific heterokaryons were obtained by polyethylene glycol induced fusion of protoplasts from auxotrophic mutants of P. ostreatus+P. cornucopiae, P. ostreatus+P. florida, P. ostreatus+P. sajor-caju, P. florida+P. cornucopiae, P. florida+P. sajor-caju and P. sajor-caju+P. cornucopiae protoplasts. The Fusion products of protoplasts were induced by MCM+benomyl, but segregation of sectors were not identified. Interspecific fusion products of protoplasts between incompatible strains did not form true clamp connections and did not produce fruit bodies. For induction of fertility, interspecific heterokaryons crossed with their parents by hyphal anastomosis.

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Establishment of Elution and Concentration Procedure for Rapid and Sensitive Detection of Norovirus in Foods of Diverse Matrices (다양한 매트릭스가 혼합된 식품을 대상으로 노로바이러스 신속검출을 위한 탈리 및 농축방법 확립)

  • Ahn, Jaehyun;Kwon, Youngwoo;Lee, Jeong-Su;Choi, Weon Sang
    • Journal of Food Hygiene and Safety
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    • v.30 no.2
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    • pp.150-158
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    • 2015
  • This article reports the development of an effective test procedure for detection of norovirus (NoV) in foods of diverse matrices. In this study, target foods included fermented milk, soybean paste, powders made from uncooked grains and vegetables, sesame leaves preserved in soy sauce, pickled mooli, and mooli. Viral recovery varied depending on the food matrices or elution buffers tested. Buffers were compared to determine effective elution buffers from artificially virus-contaminated foods. The conventional test procedure for concentrating viruses from food (elution-polyethylene glycol(PEG) precipitation-chloroform-PEG precipitation) was modified to save time by eliminating one PEG precipitation step. The modified procedure (elution-chloroform-PEG precipitation) was able to concentrate viruses more effectively than the conventional procedure. It also removed RT-PCR inhibitors effectively. The modified procedure was applied to target food for genogroup II NoV detection. NoV RNA was detected at the initial inoculum levels 3.125-12.5 RT-PCR units per 10-25 g tested food. The use of this newly established procedure should facilitate detection of low levels of norovirus in diverse foods.

Physical Properties of Mixed ${\kappa}/{\lamda}$- and ${\kappa}/{\iota}$-carrageenan Films ((${\kappa}/{\lambda}$${\kappa}/{\iota}$-carrageenan 혼합 필름의 물성)

  • Han, Yun-Jeong;Kim, Suk-Shin
    • Korean Journal of Food Science and Technology
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    • v.40 no.1
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    • pp.42-46
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    • 2008
  • The physical properties of ${\kappa}/{\lamda}$- and ${\kappa}/{\iota}$-carrageenan films, mixed with ${\lamda}$- or ${\iota}$-carrageenan (${\kappa}:{\lamda}$ or ${\kappa}:{\iota}$ = 2:1, 1.5:1.5, 1.2:1.8) and with polyethylene glycol, were compared to those of ${\kappa}$-, ${\lamda}$-, or ${\iota}$-carrageenan films. The tensile strengths of the ${\kappa}/{\lamda}$- and ${\kappa}/{\iota}$-carrageenan films decreased with increasing mixing ratios of ${\kappa}/{\lamda}$- or ${\kappa}/{\iota}$-carrageenan, respectively, and were lower than those of the ${\lamda}$- and ${\iota}$-carrageenan films. The elongations and water vapor permeabilities of the ${\kappa}/{\lamda}$- and ${\kappa}/{\iota}$-carrageenan films increased with increasing mixing ratios of ${\lamda}$- and ${\iota}$-carrageenan, respectively, and were higher than those of the ${\lamda}$- or ${\iota}$-carrageenan films.