• Journal of Plant Biotechnology
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• v.34 no.1
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• pp.31-36
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• 2007

Though higher plants car not metabolize D-amino acid, many prokaryotes and eukaryotes have the D-amino acid metabolism. Therefore, we transformed tobacco plants with D-amino acid oxidase (DAO), which can metabolize D-amino acid, and confirmed that transgenic tobacco plants might metabolize D-amino acid. Transgenic tobacco plants were survived a high concentration of D-serine, however non-transgenic plants were not grown on D-serine medium. From Southern and Northern blot analysis, transgenic tobacco plants selected on D-serine medium were confirmed by insert and expression of transgene. $T_{1}$ tobacco seeds derived $T_{0}$ tobacco plants selfing were grown on D-serine medium and showed normal phenotype compared to wild tobacco plants. Transgenic tobacco plants displayed the metabolic capability of D-serine. Therefore, we suggested that DAO is useful selectable marker gene for plant transformation.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.41-45
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• 2008

Laccase (EC 1.10.3.2) is a small group of enzymes that catalyze the oxidation of a broad range of phenolic compounds including hazardous and recalcitrant pollutants in the environment. This study attempted to develop an efficient system for production of a recombinant laccase by chloroplast genetic transformation of tobacco. Chloroplast transformation vector was constructed and introduced into the tobacco chloroplast genome using particle bombardment. Chloroplast-transformed plants were subsequently regenerated. PCR and southern blot analyses confirmed stable integration of the laccase gene into the chloroplast genome. Northern blot analysis revealed that mRNA of the laccase gene was highly expressed in chloroplast-transformed plants.

• The Plant Pathology Journal
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• v.36 no.4
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• pp.378-383
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• 2020

The objective of this research was introduction of chit42 to tuber mustard plants through Agrobacteriummediated transformation against white mold caused by Sclerotinia sclerotiorum. The binary plasmid pGisPEC1 was used in this study. Polymerase chain reaction analysis detected the transgene in 27 transformants with a transformation efficiency of 6.9%. Southern blot test was used to assess the copy number of transgene in tuber mustard plants. One, two, two, and two chit42-related bands were observed in the transformed lines TMB4, TMB7, TMB12, and TMB18, respectively. Enzymatic tests showed a significant increase in the activity of endochitinase in protein isolated from leaf tissues of chit42 transgenic 75-day tuber mustard lines. The pathogenicity of three pathogen isolates was tested on the leaves of transformed plans. The results of current study showed that expression of the gene chit42 in tuber mustard plants markedly reduced infection radius on the leaves 7 days after inoculation with the fungus.

• Journal of Plant Biotechnology
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• v.36 no.4
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• pp.366-372
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• 2009

This study was conducted to develop new lines expressing the characteristic of early flowering by introducing MdMADS2 gene in chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) ‘Zinba'. Transformation of chrysanthemum was conducted by Agrobacterium tumefaciens LBA4404 harboring the binary vector containing MdMADS2 controlled by double CaMV 35S promoters. Ninety three shoots were regenerated from 1,463 leaf segment explants cultured on the first selection medium (MS basal salts + 1.0 mg/L BA + 0.5 mg/L IAA + 10 mg/L kanamycin + 400 mg/L cefotaxime, pH 5.8) after co-cultivation, and 20 out of the 93 shoots rooted on the second selection medium containing 20 mg/L kanamycin and 400 mg/L cefotaxime. Many escapes (98.6%) were removed on the selection stage for rooting. Nineteen lines were confirmed as transgenic plant with transgene by PCR analysis. Six transgenic plants flowered 2-11 days earlier than non-transgenic plant without big change of phenotype, and especially, 3 (Mo-7, Mo-11, Mo-17) out of 6 transgenic lines showed a significant reduction in days to flowering compared to non-transgenic plant. Introduction and expression of MdMADS2 gene in them were confirmed by Southern and real-time PCR analyses, respectively.

• 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.

• Journal of Aquaculture
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• v.16 no.1
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• pp.51-58
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• 2003

The first Indian transgenic fish was generated in 1991 using borrowed constructs from foreign sources. To construct transformation vectors for the indigenous fishes, growth hormone genes of rohu (r-CH), Labeo rohita and catfish, Heteropneustes fossilis were isolated, cloned and sequenced; their fidelity was confirmed in prokaryotic and eukaryotic systems. A vector was constructed with grass carp b-actin promoter driving the expression of r-GH. Rohu eggs are large. fragile and swell 2~3 times. when fertilized. Hence they were amenable only for electroporated sperm-mediated gene transfer. Accordingly, the sperm electroporation technique was standardized to ensure 25% hatchling survival and 37% Presumptive transgenics without suffering any deformity. Southern analysis confirmed genomic integration in 15% of the tested individuals (Ti) belonging to family lines 2 and 3: another 25% of the Juveniles (Te) were also proved transgenic but with the transgene persisting extrachromosomally for longer than 1 to 2 years. perhaps due to the presence of replicon in the vector. Transgenics belonging to different family lines grew 6~8 times faster than the respective controls. Difference in growth trends of Ti and Te within a family line was not significant. In the Ti family 3 remarkable growth acceleration was sustained for a period longer than 36 weeks but in those of family 2, it gradually decreased. All transgenic fishes including the rohu converted the food at a significantly higher efficiency. Barring the transgenic mudloach, all the other transgenic fishes consumed food at significantly reduced rate.

• Molecules and Cells
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• v.19 no.1
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• pp.131-136
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• 2005

The ETV6-NTRK3 gene fusion, first identified in the chromosomal translocation in congenital fibrosarcoma, encodes a chimeric protein tyrosine kinase with potent transforming activity. ETV6-NTRK3-dependent transformation involves the joint action of NTRK3 signaling pathways, and aberrant cell cycle progression resulting from activation of Mek1 and Akt. The level of glutathione (GSH) was found to be markedly increased in ETV6-NTRK3-transformed NIH3T3 cells. The activities of the two GSH biosynthetic enzymes as well as of glutathione peroxidase, together with their mRNAs, were also higher in the transformed cells. The transformed cells were able to grow in the presence of GSH-depleting agents, whereas the control cells were not. L-Buthionine-(S,R)-sulfoximine (BSO) inhibited activation of Mek1 and Akt in the transformed NIH3T3 cells. These observations imply that up-regulation of GSH biosynthesis plays a central role in ETV6-NTRK3-induced transformation.

• Korean Journal of Microbiology
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• v.27 no.1
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• pp.21-26
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• 1989

The chromosomal DNA of Klebsiella penumoniae KNF1 was partially digested with HindIII. pBR322 was completely digested with same enzyme with additional alkaline phosphatase treatment. Both DNA samples were ligated and transformed into E. coli KO60. Single coloneis of the transformed cells were isolated on N0free agar media. These colonies were ampicillin-resistant and tetracycline-sensitive. When the plasmids in transformants were cured, nitrogen-fixing activities were lost. Therefore, these transformants harbored recombinant plasmid including nif genes of K. pneumoniae. Nitrogenase activity of transformant was higher than or the same as that of K. pneumoniae KNF1.

• Mycobiology
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• v.47 no.3
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• pp.350-354
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• 2019

In the post-genomic era, gene function analysis has attracted much attention. Transformation is often needed to investigate gene function. In this study, an easy, rapid, reliable, and cost-effective colony polymerase chain reaction (PCR) method for screening mushroom transformants was developed: picking up a suitable amount of transformant's tissue ($1-10{\mu}g$) to $20{\mu}l$ 0.25% Lywallzyme solution, and vortexing for 10 s followed by incubation at $34^{\circ}C$ for 15 min. Finally, $2{\mu}l$ of the suspension was used as templates to perform PCR and single target bands were successfully amplified from respective transformants of Tremella fuciformis, Pleurotus ostreatus, and Pleurotus tuber-regium. This procedure could be widely employed for screening transformants in mushroom transformation experiments.

• Journal of Plant Biotechnology
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• v.2 no.1
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• pp.25-28
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• 2000

The anthocyanin gene encoding flavonoid 3',5'-hydroxylase(F3,5H) was normally expressed in Nicotiana tobacco (Xanthi) plants cocultivated with Agrobacterium tumefaciens LBA4404 carrying egg plant flavonoid 3',5'-hydroxylase cDNA. Northern blot analysis showed the normal expression of F3', 5'H gene from transgenic plants. Here we found the phenotypic differences between transgenic plants and wild-type plants. The petal shape of transgenic plants showed more round shape and around petal tube area was compared to that of wild-type tobacco plants. And the petal color of transgenic plants was much lighter than that of wild-type tobacco plants.