• BMB Reports
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• v.57 no.2
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• pp.86-91
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• 2024

The fatty acids content of castor (Ricinus communis L.) seed oil is 80-90% ricinoleic acid, which is a hydroxy fatty acid (HFA). The structures and functional groups of HFAs are different from those of common fatty acids and are useful for various industrial applications. However, castor seeds contain the toxin ricin and an allergenic protein, which limit their cultivation. Accordingly, many researchers are conducting studies to enhance the production of HFAs in Arabidopsis thaliana, a model plant for oil crops. Oleate 12-hydroxylase from castor (RcFAH12), which synthesizes HFA (18:1-OH), was transformed into an Arabidopsis fae1 mutant, resulting in the CL37 line producing a maximum of 17% HFA content. In addition, castor phospholipid:diacylglycerol acyltransferase 1-2 (RcPDAT1-2), which catalyzes the production of triacylglycerol by transferring HFA from phosphatidylcholine to diacylglycerol, was transformed into the CL37 line to develop a P327 line that produces 25% HFA. In this study, we investigated changes in HFA content when endogenous Arabidopsis PDAT1 (AtPDAT1) of the P327 line was edited using the CRISPR/Cas9 technique. The successful mutation resulted in three independent lines with different mutation patterns, which were transmitted until the T4 generation. Fatty acid analysis of the seeds showed that HFA content decreased in all three mutant lines. These findings indicate that AtPDAT1 as well as RcPDAT1-2 in the P327 line are involved in transferring and increasing HFAs to triacylglycerol.

• Korean Journal of Animal Reproduction
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• v.20 no.4
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• pp.453-458
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• 1997

It is known that the incorporation of genes into transgenic mice is generally stable and is p passed on to succeeding generations in a Mendelian fashion. In this report, transgenic mice were set as a model to evaluate whether the transgenes are transmitted in a Mendelian principle in a successive generations and how they are tran s smitted into their offspring. A 3.0 kb linear DNA fragment, containing the MMTV LTR, bovine aSI casein cDNA and SV 40 splicing and polyadenylation site; was microinjected into fertilized mouse embryos. The tail DNAs of the resulting pups were subjected to dot and Southern hybridizations to screen transgenic founders. The DNAs of their offspring were anlyzed by PCR to confirm the transmission of the transgene from F0. Out of 72 live pups four pups (5.6%), 3 males and 1 female, were positive for the transgene. The rates of transmission from F0 into F1 were 33.3, 7.7, 0, and 62.5%. Those from F1 into F2 were 63.6, 5.9, and 68.8% and those from F2 into F3 were 85.7, and 88.2%. In this report, the transmission pattern of transgenes in transgenic mice into their offspring was demonstrated. It either follows or does not follow in a Mendelian fashion. Deletion or loss of the transgenes from F0 in some lines became apparant to the succeeding generations.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.540-546
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• 2010

This study was conducted to understand the role of soybean pathogenesis-related 10 (GmPR10) gene in salt tolerance and to develop salt-tolerant rice using GmPR10 cDNA. GmPR10 transgene was expressed constitutively in the shoot and root of the $T_1$ transgenic rice plants. Interestingly, however, the levels of the transgene expression were increased temporally up to over four- to five-fold in the shoot and root by 125 mM NaCl treatment, peaking at six hours after the treatment and decreasing thereafter. Electrolyte leakage of leaf cells under 125 mM NaCl treatment was lower in all the transgenic lines than in the control variety, Dongjin-byeo. Ability of seedlings to recover from 125 mM NaCl treatment for two weeks was higher in the transgenic plants than in the control plants. These results demonstrated that GmPR10 had function to increase cell integrity and promote growth under the saline stress imposed by NaCl. The transgenic line GmPR10-3 which showed highest ability to recover from the saline stress could be used as a potential source for salt tolerance in rice breeding programs.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.107-107
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• 2003

The activation of protooncogenes or the inactivation of their gene products may be a specific and effective functional study for human neoplasia. To examine this possibility, we have used the tetracycline regulatory system to generate transgenic mice that conditionally express the HccR-2 protooncogene in vivo. The new human cervical cancer protooncogene (HccR-2) was detected from cervical cancer cell line. To elucidate its biological functions, we generated transgenic mice that expressed the HccR-2 gene. The sustained expression of the HccR-2 transgene culminated chronic neutrophilic leukemia (CNL). CNL is a rare chronic myeloproliferative disorder that presents as a sustained, mature neutrophilic leukocytosis with few or no circulating immature granulocytes, the absence of peripheral blood monocytosis, basophilia, or eosinophilia, and infiltration of neutrophils at the liver, spleen and kidney. Mice expressing the HccR-2 and tetracycline-transactivating protein (tTa) transgene were found to have altered myeloid development that was characterized by increased percentages of mature neutrophil and band form neutrophil in the peripheral blood, liver and spleen. Activation of the transgene causes CNL. In our model, expression of HccR-2 transgene mice was similar in many respects to the human CNL. This model will be valuable not only for investigating the biological properties of the HccR-2 and other protooncogenes in vivo but also for analyzing the mechanism involved in the progression of CNL.

• Korean Journal of Poultry Science
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• v.26 no.2
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• pp.119-129
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• 1999

In recent years, numerous researches have been carried out in author's laboratory to develop several kinds of methods for producing transgened chicken, leaving a lot of new findings. Some of them are very useful to search for new approaches necessary to improve the efficiency of hatchability and the survival rate of developing trasgened embryos. The results obtained hitherto might be summarized as follows: (1) foreign gene(Lac Z/ Miw Z) introduced into blastodermal cells of developing embryos was successfully transferred to embryos, leading to the production of primordial germ cells(PGCs) carrying foreign DNA. However, hatched hickens failed to show the incorporation of introduced gene into the gonads. (2) When foreign gene was introduced into germinal crescent region (GCR), the gene was also efficiently incorporated into germ cells, resulting in the production of transgened chickens(offspring) which produced fruther offspring having foreign gene in the gonads. In this case, 2nd and 3rd generations of chickens were obtained through the reproduction of transgened birds. (3) In another way, the gene was injected into blood vessels of developing embryos at stage 13∼15, creating PGCs having foreign gene, and produced some transgened chickens. In this work, the PGCs were transfered between embryos, resulting in the production of transgenic chickens. (4) in these experiments, PGCs were effectively employed for producing transgenic birds, developing some kinds of chimeric chickens from homo- or hetero-sexual transfer of the PGCs from embryos. This means that the gonads from donor PGCs developed in some degree to the stage of hatching. However, these gonads showed slightly abnormal tissues similar to ovotestis like organs through histological examination. (5) Avian Leukosis Virus(ALV) induced B cell line(DT40) successfully carried foreign genes into chicken embryos, suggesting the possibility of the cells as a vector in this field of study in the future. (6) Inter-embryonic transfer of the PGCs also gave us some.

• Molecules and Cells
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• v.25 no.2
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• pp.231-241
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• 2008

Indole glucosinolates (IG) play important roles in plant defense, plant-insect interactions, and stress responses in plants. In an attempt to metabolically engineer the IG pathway flux in Chinese cabbage, three important Arabidopsis cDNAs, CYP79B2, CYP79B3, and CYP83B1, were introduced into Chinese cabbage by Agrobacterium-mediated transformation. Overexpression of CYP79B3 or CYP83B1 did not affect IG accumulation levels, and overexpression of CYP79B2 or CYP79B3 prevented the transformed callus from being regenerated, displaying the phenotype of indole-3-acetic acid (IAA) overproduction. However, when CYP83B1 was overexpressed together with CYP79B2 and/or CYP79B3, the transformed calli were regenerated into whole plants that accumulated higher levels of glucobrassicin, 4-hydroxy glucobrassicin, and 4-methoxy glucobrassicin than wild-type controls. This result suggests that the flux in Chinese cabbage is predominantly channeled into IAA biosynthesis so that coordinate expression of the two consecutive enzymes is needed to divert the flux into IG biosynthesis. With regard to IG accumulation, overexpression of all three cDNAs was no better than overexpression of the two cDNAs. The content of neoglucobrassicin remained unchanged in all transgenic plants. Although glucobrassicin was most directly affected by overexpression of the transgenes, elevated levels of the parent IG, glucobrassicin, were not always accompanied by increases in 4-hydroxy and 4-methoxy glucobrassicin. However, one transgenic line producing about 8-fold increased glucobrassicin also accumulated at least 2.5 fold more 4-hydroxy and 4-methoxy glucobrassicin. This implies that a large glucobrassicin pool exceeding some threshold level drives the flux into the side chain modification pathway. Aliphatic glucosinolate content was not affected in any of the transgenic plants.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.435-439
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• 2000

Explants of lettuce (Lactuca sativa L.) were cocultured with A. tumefaciens LBA 4404 harboring ${\gamma}$-tocopherol methyltransferase (${\gamma}$-TMT) gene from Arabidopsis thaliana. These explants were transferred to MS medium supplemented with 50 mg/L kanamycin, 500 mg/L carbenicillin, 0.1 mg/L NAA and 0.5 mg/L BA. After 4 weeks, kanamycin resistant shoots were obtained from the explants on the selection medium. The putative transgenic shoots were transferred to rooting MS medium supplemented with 50 mg/L kanamycin and 250 mg/L carbenicillin. Stable incorporation of the Arabidopsis ${\gamma}$-TMT cDNA into lettuce genomic DNA was confirmed by PCR and Southern analysis. HPLC analysis showed that $\alpha$- to ${\gamma}$-tocopherol ratio increased over four fold in a transgenic lettuce line indicating successful expression of the transgenic Arabidopsis ${\gamma}$-TMT in lettuce.

• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.351-355
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• 1995

A truncated Polygalacturonase (PG) cDNA was fused in reverse orientation to the CaMV 35S promoter of the binary vector pCA643, and introduced into tomato cells by Agrobaderium - mediated transformation. Transformed cells were selected using kanamycin as select agent then regenerated into plants. After selfed, one transgenic line (T9), was germinated and grown on MS medium containing 1 mg/mL of kanamycin Genomic Southern analysis of a T9 progeny with labelled PG2 cDNA probe showed a single antisense PC fragment as well as the endogenous PG2 gene, suggesting that PC antisense gene was integrated into tomato genome. Northern blot analysis demonstrated that the antisense RNA was produced from the transgene at much tiger level than the endogenous PG2 gene. Polygalacturonase activity analysis of the fruit from transgenic plants demonstrated that the antisense transgene expression caused 4 to 60% reduction of endogenous PG activity.

• BMB Reports
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• v.53 no.4
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• pp.229-233
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• 2020

The anti-colorectal cancer monoclonal antibody CO17-1A (mAb CO), which recognizes the tumor-associated antigen EpCAM, was expressed in transgenic Arabidopsis plants. PCR and western blot analyses showed the insertion and expression of heavy chain (HC)/HC fused to the KDEL ER retention modif (HCK) and light chain (LC) of mAb CO and mAb CO with HCK (mAb COK) in Arabidopsis transformants. Both plant-derived mAb^P CO and mAb^P COK were purified from a biomass of approximately 1,000 seedlings grown in a greenhouse. In sandwich ELISA, both mAb^P CO showed a slightly higher binding affinity for the target, EpCAM, compared to mAb^M CO. In cell ELISA, both mAbs^P COs showed binding affinity to the human colorectal cancer cell line SW480. Furthermore, mAb^M CO, mAb^P CO, and mAb^P COK exhibited dose and timedependent regression effects on SW480 cells in vitro. In summation, both mAb^P CO and mAb^P COK, expressed in Arabidopsis, recognized the target antigen EpCAM and showed anti-proliferative activity against human colorectal cancer cells.

• Journal of Life Science
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• v.14 no.4
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• pp.644-649
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• 2004

During plant development, active gibberellins (GAs) control many aspects of plant growth and development including seed germination, stem elongation, flower induction, anther development and seed growth. To understand the biosynthesis and functional role of active GAs in high plants, this study investigated GA 3$\beta$-hydroxylase gene en-coding $GA_1$ and$GA_4$ catalizing last step in GA biosynthetic pathway. The antisense GA 3$\beta$-hydroxylase gene was inserted into expression vector, pIG121-Hm. Calli derived from mature seeds of rice (Oryza satiiva L. cv. Donjinbyeo) were co-cultivated with Agrohacterium tumefaciens EHA101 earring a pIG121-Hm containing hygromycin resistance ($Hyg^r$) and antisense GA 3$\beta$-hydroxylase gene. Seventeen transgenic plants obtained inhibiting GA 3$\beta$-hydroxylase. Transgenic plants had shorter plant height more than that of the Dongjinbyeo. Stable integration of antisense GA 3$\beta$-hydroxylase gene was confirmed by polymerase chain reaction of genomic DNA isolated from the leaf organs of the $T_o$ generation.