• Korean Journal of Agricultural Science
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• v.28 no.2
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• pp.92-98
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• 2001

Lactofenicin is an antibacterial peptide fragment (about 5 kD) derived from lactoferrin (80 kD) that displays the various biological functions. The production of a human lactoferricin (Lactoferricin H) in mouse HC11 mammary epithelial cells was achieved by placing its cDNA under the control of the bovine ${\beta}$-casein gene. To express lactoferricin H in this cell culture system, constructed a hybride-splice signal consisting of bovine ${\beta}$-casein intron I and rabbit ${\beta}$-globin intron II, and a DNA fragment spanning intron 8 of the bovine ${\beta}$-casein gene. Expression of lactofenicin H from this expression vector was identified by RT-PCR, northern and dot blot analysis. RT-PCR using total RNA of HC11 cells transfected with pBL1-cin expression vector yielded a product identified as having a size of the 150bp. Northern blot analysis was identified about 2.3 kb. In dot blot analysis, recombinant lactofenicin H was recognized with anti-human lactofrrnin polyclonal antibody.

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.87-90
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• 2001

BcGRl gene encoding cytosolic glutathione reductase of Chinese cabbage (Brassica campestris var. Pekinensis cv. Seoul) was placed under the control of the CaMV 35S promoter and introduced into tobacco (Nicotiana tabacum L. cv. Samsun) via Agrobacterium-mediated transformation. T$_{0}$ 32 independent plants transformed with BcGRl gene were selected with kanamycin and they were confirmed by polymerase chain reaction (PCR) and Southern blot analysis. Northern blot analysis revealed that the constitutive expression of BcGRl gene and there was no relationship between the copy number of introduced gene and the levels of BcGRl transcripts.

• Journal of The Korean Society of Grassland and Forage Science
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• v.21 no.1
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• pp.21-26
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• 2001

To develop transgenic orchardgrass resistant to reactive oxygen species produced from environmental stresses, a vector with the cytosolic glutathione reductase cDNA (BcGRl) from Chinese cabbage was constructed under the control of the cauliflower mosaic virus 35S promoter and was introduced into orchardgrass using Agrobacterium tumefaciens EHA101. Transgenic plants from hygromycin-selected calli of orchardgrass did not show any morphological difference from wild-type plants. The results of PCR amplification and genomic Southern blot analysis confirmed the integration of foreign gene into the chromosome of transgenic orchardgrass. Northern blot analysis with total RNA from leaves also confirmed the constitutive expression of BcGR1 in transgenic orchardgrass.

• Korean Journal of Animal Reproduction
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• v.18 no.2
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• pp.133-139
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• 1994

Two human lactoferrin expression vectors(pCChcLf and pCChcLf-1) were constructed using rat $\beta$-casein gene and human lactoferrin cDNA. The recombinant DNAs containing human lactoferrin cDNA were microinjected into the fertilized eggs of hybrid mice (BDF1 : C57BL$\times$DBA) and the DNA-injected eggs were treansferred into the oviducts of foster mothers. Genomic DNAs were isolated from the tails of mice born from the microinjected eggs and analyzed by Southern blot analysis. As a result, 5 and 9 transgenic mice with CChcLf and CChcLf-1 gene were produced, respectively. To determine tissue-specificity of transgene expression, Northern blot analysis was performed. Female transgenic mice were killed at day 10 of lactation and total RNAs from various tissues were isolated. Based on Northern blot analysis, it was shown that transgene was mainly expressed in the mammary glands of transgenic mice. In addition, the human lactoferrin in milk was detected by enzyme-linked immunosorbent assay. For this study, milk was obtained from the mammary glands of the transgenic mice at day 10 of lactation. In line #2 of CChcLf and line #7 of CChcLf-1 transgenic mice, human lactoferrin was secreted into the milk at concentration levels of 340ng/ml and 60ng/ml, respectively.

• Journal of Plant Biotechnology
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• v.7 no.1
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• pp.45-50
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• 2005

To clone blue and white flower color specific genes, mRNA differential display was carried out with two different Commelina species, C. communis Linne for blue color and C. coreana Leveille for. leucantha Nakai for white color. Fifty two and 100 cDNA clones specific for blue or white flower color, respectively, were ranging from 200 to 700 bp in size. From the reverse northern blot analysis, 12 and 7 positive clones were selected for blue and white flower, respectively. These clones appear to be novel cDNAs for these Commelina plants, but not color specific. This finding was supported by the northern blot analysis. However, two clones, B18 and B19, derived from blue flowered Commelina were highly expressed than in the white Commelina species, implying that further study will be valuable. The results indicated that both mRNA display experiment and dot blot analysis may not sensitive enough to clone color-determining gene from the plant, leading to explore more advanced method, like high-density colony array study (HDCA).

• Journal of Plant Biotechnology
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• v.1 no.2
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• pp.85-90
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• 1999

In order to protect fungal diseases, leaf disc explants of Solanum tuberosum cultivar, Belchip, was infected with an Agrobacterium MP90 strain containing chimeric gene construct, consisting of antibiotic resistance and chitinase gene driven by the CaMV 35S promoter, for transformation. Regenerated multiple shoots were selected on a medium containing kanamycin and carbenicillin after exposure to Agrobacterium. The presence and integration of the npt II and chitinase gene were confirmed by polymerase chain reaction(PCR). Northern blot analysis indicated that the genes coding for the enzyme could be expressed in potato plants. The chitinase activity of transgenic potato plants was higher than the control potato.

• Korean Journal of Plant Resources
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• v.19 no.3
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• pp.440-446
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• 2006

To investigate flowering related genes in winter-type oilseed rape (Brassica napus L. cv. Tammi), differentially expressed genes were isolated from leaves of the plant after low temperature treatment which is requirements for floral induction. As a result of suppression subtractive hybridization (SSH), 288 clones were randomly selected from SSH library. Using reverse Northern blot analysis, 150 of 288 clones were identified to be differentially expressed. Out of these 150 clones, 45 clones showed very high identities with the known genes. Four clones showed very high identities over 90% with metallothionein-like gene that is related to flowering-induced genes. Of these 4 clones, the cDNA clone, rfs-13, revealed high identity with meotallothionein-like protein in Arabidopsis thaliana (98%) and Brassica compestris (89%). Furthermore, gene expressed in immature flower stages was confirmed by Northern blot analysis.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.3
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• pp.315-320
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• 1999

Molecular regulation of the renin-angiotensin system (RAS) was investigated in deoxycorticosterone acetate (DOCA)-salt hypertension. The expression of renin, angiotensinogen and angiotensin II receptor genes in the kidney and liver was determined by Northern blot analysis in rats which were made DOCA-salt hypertensive over the period of 2 or 4 weeks. Along with the hypertension, renin mRNA was decreased in the remnant kidney. The expression of angiotensinogen gene was not significantly altered in the kidney, but was significantly decreased in the liver. The expression of angiotensin II receptor gene was increased in the kidney, while it remained unaltered in the liver. The duration of hypertension did not affect the altered gene expression. It is suggested that the components of RAS are transcriptionally regulated in DOCA-salt hypertension in a tissue-specific manner.

• Journal of Plant Biotechnology
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• v.40 no.4
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• pp.198-202
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• 2013

To produce transgenic cucumber expressing Nit gene coffering abiotic resistance, the cotyledonary-node explants of cucumber (cv. Eunsung) were inoculated with A. tumefaciens transformed with pPZP211 or pCAMBIA2300 carrying Nit gene, that has cis-acting element involved in resistance to various abiotic environmental stresses. After co-cultivation, the procedures of selection, shoot initiation, shoot elongation, and plant regeneration were followed by cotyledonary-node transformation method (CTM, Jang et al. 2011). The putative transgenic plants were selected when shoots were grown to a length greater than 3 cm from the cotyledonary-node explants on selection medium supplemented with 100 mg/L paromomycin as a selectable agent. The confirmation of transgenic cucumber was based on the genomic PCR, Southern blot analysis, RT-PCR, and Northern blot analysis. A 105 shoots (4.12%) selected from the selection mediums were obtained from 2,547 explants inoculated. Of them, putative transgenic plants were only confirmed with 45 plants (1.77%) by genomic PCR analysis. Transgenic plants showed that the Nit genes integrated into each genome of 39 plants (1.53%) by Southern blot analysis, and the expression of gene integrated into cucumber genome was only confirmed at 6 plants (0.24%) by RT-PCR and Northern blot analysis. These results lead us to speculate that the genes were successfully integrated and expressed in each genome of transgenic cucumber.

• Plant Biotechnology Reports
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• v.3 no.4
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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.