• 한국초지조사료학회지
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• 제20권2호
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• pp.103-108
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• 2000

To produce of transgenic orchardgrass, the seed-derived calli of orchardgrass (Dactylis glomerata L.) co-cultivated with Agrobacterium turnefaciens EHAlOl harboring binary vector pIG121-Hm were selected with hygromycin and then transferred onto N6 regeneration medium containing 1 rngl l of NAA, 5 rngl l of kinetin, 250 rngl l of carbenicillin and 50 mg/ l of hygromycin. The efficiency of transformation was differed on cultivars, that is, 'Potomac' appeared 12% of transformation efficiency while 'Amba' did 5.5%. The addition of acetosyringone during co-cultivation was a key to successhl transformation of orchardgrass. Transgene fragments were identified by PCR analysis and the constitutive expression of GUS gene was confirmed by Northern blot analysis. (Key words : Acetosyringone, Agrobacterium tumefaciens, Orchardgrass (Dactylis glomerata L.), Transformation)

• 한국항만경제학회지
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• 제21권3호
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• pp.239-258
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• 2005

Port e-Transformation would employ new technologies such as CRM, SCM, ERP to all the port facilities and improve the efficiency of management for ports. In this paper, firstly, we analyze classification organization of Port e-Transformation technology. Secondly, understand the current situation on each component of Port e-Transformation. Thirdly, present tasks for each component of it. Finally, propose problems and its solving methods for these tasks. However, rationalization of management for internal structure occurred during process of Port e-Transformation and new adjustment of laws related to port are not included in this paper.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.294-301
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• 2001

The effects of including glycine and isonicotinic acid hydrazide (INH) in the growth medium (Luria broth, LBG) on the subsequent lysozyme-imduced protoplast formation and transformation efficiency of Corynebacterium glutamicum were studied. The transformation efficiency of C. glutamicum AS019 increased up to 100-fold as the ocncentrationof glycine in the media increased from 0% to 5% (w/v), relative to cells grown in the absence of glycine. The presence of 5 mg/ml INH in the growth medium led to a further 10-fold increase in transformation efficiency. In addition, this transformation protocol was successfully applied to other strains of C. glutamicum. Both chemicals affected the mycolic acid attachment to the cell surface of C. glutamicum, when INH, the relative percentage of fatty acids of AS019 to the total lipids (mycolic acid plus fatty acids) decreased from 76.9% (in LBG) to 72.9% (in LBG-2% glycine) and 66.4% (in LBG-8 mg InG/ml), thereby suggeting that these chemicals also inhibit fatty acid synthesis.

• Journal of Microbiology and Biotechnology
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• 제22권9호
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• pp.1279-1287
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• 2012

We established an efficient transformation method for thermophile Geobacillus kaustophilus HTA426 using conjugative transfer from Escherichia coli of host-mimicking plasmids that imitate DNA methylation of strain HTA426 to circumvent its DNA restriction barriers. Two conjugative plasmids, pSTE33T and pUCG18T, capable of shuttling between E. coli and Geobacillus spp., were constructed. The plasmids were first introduced into E. coli BR408, which expressed one inherent DNA methylase gene (dam) and two heterologous methylase genes from strain HTA426 (GK1380-GK1381 and GK0343-GK0344). The plasmids were then directly transferred from E. coli cells to strain HTA426 by conjugative transfer using pUB307 or pRK2013 as a helper plasmid. pUCG18T was introduced very efficiently (transfer efficiency, $10^{-5}-10^{-3}\;recipient^{-1}$). pSTE33T showed lower efficiency ($10^{-7}-10^{-6}\;recipient^{-1}$) but had a high copy number and high segregational stability. Methylase genes in the donor substantially affected the transfer efficiency, demonstrating that the host-mimicking strategy contributes to efficient transformation. The transformation method, along with the two distinguishing plasmids, increases the potential of G. kaustophilus HTA426 as a thermophilic host to be used in various applications and as a model for biological studies of this genus. Our results also demonstrate that conjugative transfer is a promising approach for introducing exogenous DNA into thermophiles.

• Journal of Plant Biotechnology
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• 제46권4호
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• pp.323-330
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• 2019

The high expression level of industrial and metabolically important proteins in plants can be achieved by plastid transformation. The CaIA vector, a Capsicum-specific vector harboring aadA (spectinomycin resistance), is a selectable marker controlled by the PsbA promoter, and the terminator is flanked by the trnA and trnI regions of the inverted repeat (IR) region of the plastid. The CaIA vector can introduce foreign genes into the IR region of the plastid genome. The biolistic method was used for chloroplast transformation in Scoparia dulcis with leaf explants followed by antibiotic selection on regeneration medium. Transplastomes were successfully screened, and the transformation efficiency of 3 transgenic lines from 25 bombarded leaf explants was determined. Transplastomic lines were evaluated by PCR and Southern blotting for the confirmation of aadA insertion and its integration into the chloroplast genome. Seeds collected from transplastomes were analyzed on spectinomycin medium with wild types to determine genetic stability. The increased chloroplast transformation efficiency (3 transplastomic lines from 25 bombarded explants) would be useful for expressing therapeutically and industrially important genes in Scoparia dulcis L.

• Journal of Dairy Science and Biotechnology
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• 제41권1호
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• pp.1-8
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• 2023

Response surface methodology (RSM) is a statistical approach widely used in food processing to optimize the formulation, processing conditions, and quality of food products. The homogenization process is achieved by subjecting milk to high pressure, which breaks down fat globules and disperses fat more evenly throughout milk. This study focuses on an application of RSM including the logit transformation to predict the efficiency of milk homogenization, which can be maximized by minimizing the relative difference in fat percentage between the top part and the remainder of milk. To avoid a negative predicted value of the minimum of this proportion, the logit transformation is used to turn the proportion into the logit, whose possible values are real numbers. Then, the logit values are modeled and optimized. Subsequently, the logistic transformation is used to turn the predicted logit into the predicted proportion. From our model, the optimum condition for the maximized efficiency of milk homogenization was predicted as the combination of a homogenizer pressure of 30 MPa, a storage temperature of 10℃, and a storage period of 10 days. Additionally, with a combination of a homogenizer pressure of 30 MPa, a storage temperature of 10℃, and a storage period of 50 days, the level of milk homogenization was predicted to be acceptable, even with the problem of extrapolation taken into account.

• Plant Biotechnology Reports
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• 제3권4호
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• pp.277-283
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• 2009

One of the limitations to conducting maize Agrobacterium-mediated transformation using explants of immature zygotic embryos routinely is the availability of the explants. To produce immature embryos routinely and continuously requires a well-equipped greenhouse and laborious artificial pollination. To overcome this limitation, an Agrobacterium-mediated transformation system using explants of type II embryogenic calli was developed. Once the type II embryogenic calli are produced, they can be subcultured and/or proliferated conveniently. The objectives of this study were to demonstrate a stable Agrobacterium-mediated transformation of maize using explants of type II embryonic calli and to evaluate the efficiency of the protocol in order to develop herbicide-resistant maize. The type II embryogenic calli were inoculated with Agrobacterium tumefaciens strain C58C1 carrying binary vector pTF102, and then were subsequently cultured on the following media: co-cultivation medium for 1 day, delay medium for 7 days, selection medium for $4{\times}14$ days, regeneration medium, and finally on germination medium. The T-DNA of the vector carried two cassettes (Ubi promoter-EPSPs ORF-nos and 35S promoter-bar ORF-nos). The EPSPs conferred resistance to glyphosate and bar conferred resistance to phosphinothricin. The confirmation of stable transformation and the efficiency of transformation was based on the resistance to phosphinothricin indicated by the growth of putative transgenic calli on selection medium amended with $4mg\;1^{-1}$ phosphinothricin, northern blot analysis of bar gene, and leaf painting assay for detection of bar gene-based herbicide resistance. Northern blot analysis and leaf painting assay confirmed the expression of bar transgenes in the $R_1$ generation. The average transformation efficiency was 0.60%. Based on northern blot analysis and leaf painting assay, line 31 was selected as an elite line of maize resistant to herbicide.

• Journal of Plant Biotechnology
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• 제30권3호
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• pp.215-219
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• 2003

This study was conducted to find optimum bacterial density for improving the efficiency of transformation mediated by Agrobacterium tumefaciens in apples. Regeneration(15％) and transformation frequency(10％) were increased in resuspension-culture density $A_{600}$ 1.3 from preculture density $A_{600}$ 0.7 of Agrobacterium tumefaciens in ′Fuji′. In ′Gala′, 20% regeneration and 16% transformation frequency were observed at optimum bacterial density $A_{600}$ 0.7 form preculture density $A_{600}$ 1.3. ′Mclntosh as well as "Gala" were 25％regeneration and 10％ transformation frequency. Hence a frequency optimum condition of bacterial density for the efficient transformation of apple could be depend on apple genotypes.

• Journal of Plant Biotechnology
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• 제30권1호
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• pp.27-33
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• 2003

국내에서 육종된 감자 품종들에 대하여 재분화 및 형질전환효율에 영향을 미치는 여러 요인들을 조사하고 품종별 최적 조건을 규명하고자 하였다. 최근 육종된 5품종 '추백', '남서', '자심', '조풍'과 '조원'의 잎과 줄기 조직을 재료로 사용하여 배지 내 호르몬 조성별 신초형성률을 조사하였다. 신초형성률은 잎 조직의 경우 '조원'이 GA₃0.1 mg/L, Zeatin 2.0mg/L과 NAA 0.01 mg/L가 포함된 M5 배지에서 90%이상으로 가장 높은 반면 다른 공시품종은 20% 미만이었으며 줄기조직을 치상한 경우 '조풍'이 M5 배지에서 70%의 신초형성률을 나타내었으나. 다른 품종은 25% 이하의 신초형성률을 나타내어 품종별, 조직별로 큰 차이를 나타내었다. 감자의 형질전환 시 Agrobacterium 접종과정에서 75 μM의 acetosyringone을 처리한 경우 잎 조직은 '조원'이 87.9%, 줄기 조직은 '조풍'이 68.4%로 높은 형질전환효율을 나타내어 acetosyringone을 처리하지 않은 대조구에 비하여 1.5∼4.0배의 높은 형질전환효율을 나타냈다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권10호
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• pp.1390-1394
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• 2004

We have achieved efficient transformation system for forage-type tall fescue plants by Agrobacterium tumefaciens. Mature seed-derived embryogenic calli were infected and co-cultivated with each of three A. tumefaciens strains, all of which harbored a standard binary vector pIG121Hm encoding the neomycin phosphotransferase II (NPTII), hygromycin phosphotransferase (HPT) and intron-containing $\beta$-glucuronidase (intron-GUS) genes in the T-DNA region. Transformation efficiency was influenced by the A. tumefaciens strain, addition of the phenolic compound acetosyringone and duration of vacuum treatment. Of the three A. tumefaciens strains tested, EHA101/pIG121Hm was found to be most effective followed by GV3101/pIG121Hm and LBA4404/pIG121Hm for transient GUS expression after 3 days co-cultivation. Inclusion of 100 $\mu$M acetosyringone in both the inoculation and co-cultivation media lead to an improvement in transient GUS expression observed in targeted calli. Vacuum treatment during infection of calli with A. tumefaciens strains increased transformation efficiency. The highest stable transformation efficiency of transgenic plants was obtained when mature seed-derived calli infected with A. tumefaciens EHA101/pIG121Hm in the presence of 100 $\mu$M acetosyringone and vacuum treatment for 30 min. Southern blot analysis indicated integration of the transgene into the genome of tall fescue. The transformation system developed in this study would be useful for Agrobacterium-mediated genetic transformation of tall fescue plants with genes of agronomic importance.