• Title/Summary/Keyword: K-L transformation

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Introduction and Expression of a Thaumatin-like Protein from Rice in American Ginseng Following Agrobacterium-mediated Transformation

  • Chen, W.P.;Punja, Z.K.
    • Journal of Ginseng Research
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    • v.27 no.1
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    • pp.17-23
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    • 2003
  • Agrobacterium-mediated transformation of American ginseng (Panax quinquefolius L.) with strain LBA 4404 containing a rice thaumatin-like protein gene is described. The selectable markers used were phosphinothricin acetyltransferase and hygromycin phosphotransferase genes. Epicotyl explants from seedlings were precultured for 5-7 days on Murashige and Skoog medium with ${\alpha}$-naphthaleneacetic acid and 2,4 dichlorophenoxyacetic acid at 10 ${\mu}$M and 9 ${\mu}$M, respectively (ND medium), prior to Agrobacterium infection. The explants were immersed in a bacterial suspension for 20 min. A post-infection co-culture period of 3-4 days was provided on ND medium. Selection for transformed calli was conducted on ND medium with 20 mg/L phosphinothricin followed by 100 mg/L hygromycin over an 8-month period. it transformation frequency of 24.8% was achieved at the callusing phase. The presence of the transgenes in calli was confirmed by Southern hybridization and polymerase chain reaction analysis. The expression of the thaumatin-like protein gene in ginseng calli was demonstrated by Western blot analysis. Somatic embryos were produced from both transgenic calli and suspension cultures, and plantlets were recovered that expressed the transgenic thaumatin-like protein gene.

Production of Transgenic Plants in Brassica napus Winter Cultivar 'Youngsan' (영산 유채를 이용한 형질전환체 생산)

  • Roh, Kyung-Hee;Kwak, Bo-Kyoung;Kim, Hyun-Uk;Lee, Kyeong-Ryeol;Kim, Sun-Hee;Suh, Mi-Chung;Kim, Hyo-Jin;Kim, Jong-Beom
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.26-32
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    • 2011
  • To improve genetic transformation of Brassica napus winter cultivar 'Youngsan', factors influencing shoot regeneration and transformation from cotyledonary petioles were investigated. Shoot induction was enhanced in the combination of 0.5 mg/L NAA and 2~4 mg/L kinetin. Silver nitrate was essential for successful shoot regeneration, ranging from 5 to 9 mg/L. The addition of $GA_3$ promoted plant regeneration. Among the tested Agrobacterium strains, co-cultivation times, and antibiotic selection regimes, choice of appropriate Agrobacterium strain was the most critical factor for efficient transformation of B. napus cv. 'Youngsan'. The EHA105 succinamopine strain was the most efficient and the maximum transformation efficiency was 26.8%. Transgenic shoots were selected on 10 mg/L phosphinothricin (PPT) containing media. The transgenic plants expressing bar and gus genes were resistant for commercial herbicide "Basta" and stained with X-Gluc. Southern blot hybridization indicated that the presence of one to three gus gene copies per genome and inheritance of the gus gene into the $T_1$ generation.

Efficient transformation of Actinidia arguta by reducing the strength of basal salts in the medium to alleviate callus browning

  • Han, Meili;Gleave, Andrew P.;Wang, Tianchi
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.129-138
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    • 2010
  • An efficient transformation system for high-throughput functional genomic studies of kiwifruit has been developed to overcome the problem of necrosis in Actinidia arguta explants. The system uses Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pART27-10 to inoculate leaf strips. The vector contains neomycin phosphotransferase (nptII) and ${\beta}$-glucuronidase (GUS) (uidA) genes. A range of light intensities and different strengths of Murashige and Skoog (MS) basal salt media was used to overcome the problem of browning and/or necrosis of explants and calli. Callus browning was significantly reduced, resulting in regenerated adventitious shoots when the MS basal salt concentration in the culture medium was reduced to half-strength at low light intensity ($3.4\;{\mu}mol\;m^{-2}\;s^{-1}$) conditions. Inoculated leaf strips produced putative transformed shoots of Actinidia arguta on half-MS basal salt medium supplemented with 3.0 $mg\;l^{-1}$ zeatin, 0.5 $mg\;l^{-1}$ 6-benzyladenine, 0.05 $mg\;l^{-1}$ naphthalene acetic acid, 150 $mg\;l^{-1}$ kanamycin and 300 $mg\;l^{-1}$ $Timentin^{(R)}$. All regenerated plantlets were deemed putativ transgenic by histochemical GUS assay and polymerase chain-reaction analysis.

Optimized Protocols for Efficient Plant Regeneration and Gene Transfer in Pepper (Capsicum annuum L.)

  • Mihalka, Virag;Fari, Miklos;Szasz, Attila;Balazs, Ervin;Nagy, Istvan
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.143-149
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    • 2000
  • An Efficient in vitro regeneration system and an optimized Agrobacterium mediated transformation protocol are described, based on the use of young seedling cotyledons of Capsicum annuum L. Optimal regeneration efficiency can be obtained by cultivating cotyledon explants on media containing 4 mg/L benzyladenine and 0.1 mg/L indolacetic acid. The effect of antibiotics used to eliminate Agrobacteria, as well as the toxic level of some generally used selection agents (kanamycin, geneticin, hygromycin, phosphinotricin and methotrexate) in regenerating pepper tissues were determined. To enable the comparison of different selection markers in identical vector background, a set of binary vectors containing the marker genes for NPTII, HPT, DHFR and BAR respectively, as well as the CaMV 35S promoter/enhancer-GUS chimaeric gene was constructed and introduced into four different Agrobacterium host strains.

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Optimization of the Transformation of D-Glucose to Vitamin C (Glucose를 비타민 C로 변형시키는 과정의 최적화에 대한 연구)

  • Chung, Jong-Kyeong;Goo, Yang-Mo;Kim, Kong-Hwan
    • YAKHAK HOEJI
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    • v.32 no.6
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    • pp.386-393
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    • 1988
  • Chemical transformation of D-glucose to 2-keto-L-gulonic acid and L-ascorbic acid has been examined. D-Sorbitol obtained from D-glucose was microbiologically oxidized to L-sorbose by G. suboxydans in 90% yield. On treatment of L-sorbose with acetone in the presence of sulfuric acid, its diacetonide is obtained in 95% yield. This diacetonide is oxidized to the corresponding acid with nickel chloride-hypochlorite, and the acid is directly transformed to L-ascorbic acid. The over all yield of Vitamin C from D-glucose achieved is 54%.

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Oxidative Transformation of Tetracycline in Aqueous Solution by Birnessite (망간산화물(birnessite)에 의한 수용액 중 Tetracycline의 산화-변환반응에 대한 연구)

  • Eom, Won-Suk;Kim, So-Hui;Shin, Hyun-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.37 no.2
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    • pp.73-80
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    • 2015
  • An investigation on the removals of tetracycline (TTC), which is a family of antibiotics widely founded in the environment, from the aqueous solution by birnessite(${\delta}-MnO_2$)-mediated oxidative transformation was described. This study also examined the potential effect of the naturally occurring substances, humic acid (HA) on the oxidative transformation. The experiment was carried out in various conditions (reaction time, Mn oxide loadings, pH) and in the presence of HA as a batch test. The removals of TTC followed pseudo-first order reactions, and rate constants (k, $hr^{-1}$) for the removals of TTC were constantly increased with decreasing pH from 0.98 (pH 9) to 2.97 (pH 3). The rate constants also increased about 1.3 times when the birnessite loading increased from 1 to 2 g/L. Presence of HA (5 mg-C/L, at $pH{\geq}6$) caused some enhancement in the removals of TTC as compared to the control, and also showed the removal efficiencies of TTC in the birnessite mediated systems (TTC=0.25 mM, ${\delta}-MnO_2=2.0g/L$, pH 6) increased with increasing HA concentrations (1~10 mg-C/L). The results obtained from the oxidative transformation of TTC and the effect of HA were discussed in terms of reaction characteristics and mechanism.

Effects of Antibiotics on Suppression of Agrobacterium tumefaciens and Plant Regeneration from Wheat Embryo

  • Han, Si-Nae;Oh, Poo-Reum;Kim, Hong-Sig;Heo, Hwa-Young;Moon, Jun-Cheol;Lee, Sang-Kyu;Kim, Kyung-Hee;Seo, Yong-Weon;Lee, Byung-Moo
    • Journal of Crop Science and Biotechnology
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    • v.10 no.2
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    • pp.92-97
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    • 2007
  • Antibiotics used for suppressing Agrobacterium in plant transformation procedure might have negligible effects on plant tissues and regeneration. The effects of antibiotics on growth suppression of Agrobacterium and plant regeneration were investigated for enhancing Agrobacterium-mediated transformation using wheat mature embryos. Antibiotics tested, except carbenicillin, were able to suppress that embryos were coated with a layer of Agrobacterium cells in callus induction medium. Agrobacterium growth was suppressed minimally at 50 mg/l of timentin, while cefotaxime and clavamox were completely suppressed at relative high concentration of 250 mg/l. In the treatment of carbenicillin, initiation of growth suppression of Agrobacterium occurred at 750 mg/l of concentration because Agrobacterium KYRT1 contains the carbenicillin resistant gene. In Agrobacterium inoculation, effects of antibiotics were significantly different on the rate of callus induction and shoot formation. Almost embryos were induced calli at 50 mg/l of timentin whereas callus induction rate was achieved above 90% at 100 mg/l and 250 mg/l of cefotaxime and clavamox, respectively. Shoot formation rate was higher in the treatment of timentin than that of cefotaxime and clavamox at 500 mg/l of concentration, respectively. Timentin can be used as a good antibiotics in Agrobacterium-mediated wheat transformation.

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Agrobacterium Mediated Transformation from Callus Pretreated with Particle Bombardment in Lilium lancifolium Thunb. (Particle Bombardment에 의해 전처리 된 참나리(Lilium lancifolium Thunb.) 캘러스의 Agrobacterium tumefaciens을 통한 형질전환)

  • Nam, Sang-Wook;Kim, Hei-Young
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.13-17
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    • 2004
  • To improve transformation efficiency, the callus of Lilium lancifolium Thunb. were bombarded by particles coated with pIG 121 Hm which include NPT II and GUS genes, and then cocultivated with Agrobacterium tumefaciens EHA101 which contain pIG121Hm binary vector, carrying neomycin phosphotransferase (NPT II) and $\beta$-Glucuronidase (GUS) genes. Three days after cocultivation with Agrobacterium tumefaciens and particle bombardment, the callus clusters were transferred to MS medium containing 1mg/L 2,4-D, 0.1mg/L BAP, 100mg/L kanamycin and 200mg/L carbenicillin. Four weeks after transfer to the selection medium, GUS expression was detected and PCR analysis revealed the presence of NPT II fragment of the expected size (700 bp) in the transformed callus. The GUS expression from Agrobacterium-mediated transformants after particle bombardment increased to over 3-folds compared with that of callus cocultivated with Agrobacterium tumefaciens without particle bombardment. The callus clusters containing kanamycin resistant gene were transferred to MS medium containing 1mg/L NAA and 1mg/L BAP. Somatic embryos were developed in four weeks and microbulbs expressing GUS were formed.

Effect of Caffeine on Transformation Induced by Adenovirus type 12 (Adenovirus type 12에 의해 유발된 Transformation에 미치는 Caffeine의 영향)

  • Choi, Sung-Bae
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.269-272
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    • 1996
  • Adenovirus group consists of over 100 related viruses which have been isolated from respiratory or gastro-intestinal tract of primate, cattle, dog and mice. Approximately 40 serologic types of adenovirus producing a variety of human respiratory and conjunctival infections were identified. Adenoviruses are icosahedral virions containing double-stranded linea DNA. They are 70nm to 90nm in diameter and each of capsid is composed of 252 capsomeres. Several numbers of this group, including types commonly associated with respiratory disease in man, are capable of producing malignant tumors in young hamsters and a few types have been shown to be on-cogenic in young rat. Previous report involving effect of Hormone on replication of adenovirus(9) has been carried out. The present report represents a continuation of previous study. To obtain evidence concerning the effect of caffeine on the transformation, investigation of adenovirus type 12 of this group was undertaken. For practical consideration it was desirable to investigation of the effect of caffeine on the adenovirus type 12-induced transformation in L cell. Results were as follows; 1. Adenovirus type 12-induced transformation was inhibited in the presence of caffeine. 2. Yields of adeoovirus type 12 in L cell were slightly inhibited by treatment of caffeine.

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Current status of tissue culture and genetic transformation systems in oilseed rape plants (Brassica napus L.) (유채 조직배양 및 형질전환 연구동향)

  • Lee, Sang-Il;Kim, Yun-Hye;Lee, Dong-Hee;Lee, Yu-Mi;Park, Seo-Jun;Kim, Jong-Bo
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.379-387
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    • 2010
  • Oilseed rape (Brassica napus L.) is an important crop due to its high oil content in the seed. Recently, the demand for the improvement of crop for biodisel energy source is increased as oil prices in the world has increased dramatically. Until now, oilseed rape breeding was carried out by cross-hybridization between different varieties and related germplasms. However, like as many other crops, the application of tissue culture and gene transformation systems has been introduced into oilseed rape breeding program including the development of transgenic canola plants. In this study, we reviewed a history of tissue culture and genetic transformation research in oilseed rape plants and indicated some important aspects for the production of transgenic oilseed rape plants.