• 한국육종학회지
• /
• 제40권2호
• /
• pp.128-135
• /
• 2008

Herbicide resistance is the most common trait being tested and thus herbicide?resistant genetically modified plants are now the most widely cultivated worldwide. Here we developed herbicide?resistant transgenic Agrostis mongolica Roshev. by employing an efficient Agrobacterium?mediated transformation procedure with 25.2% of transformation efficiency. The identification and employment of regenerable and reproducible type of callus was one of the most critical factors to ensure success in this study. PCR analysis confirmed that the bar transgene was integrated into the genome of transgenic plants. The expression of 35S?bar gene was confirmed by Northern blot analysis. The transgenic plants showed complete resistance to herbicide, indicating that the bar gene is functional in transgenic plants.

• Fisheries and Aquatic Sciences
• /
• 제18권1호
• /
• pp.65-71
• /
• 2015

The functional utility of a transgenic marine medaka Oryzias dancena strain carrying the red fluorescent protein (RFP) gene driven by an endogenous choriogenin H (chgH) promoter was evaluated for its ability to detect waterborne $17{\alpha}$-ethinylestradiol (EE2), a synthetic estrogen derivative. The chgH:rfp transgenic marine medaka larvae showed an age-dependent tendency in the efficiency of EE2-mediated transgene expression, in which transgenic larvae older than 6 days post-hatching displayed a more effective response in their transgene expression to EE2 than did younger hatchlings. During experimental exposures to high concentrations of EE2 (200 to 1,000 ng/L), the transgenic responses in the hatchlings were broadly dose- and duration-dependent. With exposures using lower doses of EE2 (25, 50 and 100 ng/L), EE2-induced transgenic RFP was also observed in the transgenic larvae, although the lower doses required exposure of longer duration. Under the EE2 exposure and microscope assay conditions used in our study, transgenic marine medaka larvae exhibited a similar degree of EE2-mediated RFP phenotype expression at various salinity levels (0, 15 and 30 ppt).

• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2000년도 국제심포지움
• /
• pp.7-8
• /
• 2000

Transgenic animals are very useful for scientific, pharmaceutical, and agricultural purposes. In livestock, transgenic technology has been used forthe genetic alteration of farm animals, the production of human proteins inlarge quantities in the milk of transgenic farm animals, and the generation of animals with organs suitable for xenotransplantation. To date, the transfer of foreign genes into farm animals has been performed mainly by microinjection of DNA into the pronuclei of fertilized eggs. However, the overall success rate of transgenic animals in livestock so far has been disappointingly low, eg., the efficiency is 0∼5% in swine, and less than 1% in sheep and cattle, compared with the rate in mice where 5% microinjected develop into transgenic animals. Recently, McGreath et al. (2000) have succeeded in producing the gene targeted sheep by the use of nuclear transfer from cultured somatic cells transfected with a foreign gene in vitro. However, we may need plenty of time until currently employ this method for gene transfer to farm animals. We have been studying to exploit the method for improving production efficiencies of transgenic animals with emphasis of its application to farm animals. The present paper describes three approaches that we have made in our laboratory to improve production efficiencies of transgenic animals, based on the DNA microinjection method. 1. Co-injection of restriction enzyme with foreign DNA into the pronucleus for elevating production efficiencies of transgenic animals. 2. Efficient selection of transgenic mouse embryos using EGFP as a marker gene. 3. Phenotypes of tansgenic mice expressing WAP/hGH-CAG/EGFP fusion gene produced by selecting transgenic embryos. 4. Efficient site-specific integration of the transgene targeting an endogenous lox like site in early mouse embryos.

• Journal of Crop Science and Biotechnology
• /
• 제11권2호
• /
• pp.107-110
• /
• 2008

A high-efficiency, reproducible somatic embryogenesis system for strawberry cultivar Chandler was developed. Thirty-one somatic embryos per explant(max no.) were recorded in leaf discs which were cultured on medium containing MS salts+$B_5$ vitamins+2% glucose+4.0 mg $1^{-1}$TDZ(Thidiazuron) and incubated at $10{\pm}1^{\circ}C$ under darkness for one week followed by three weeks under 16-h photoperiod. The scanning electron microscopic(SEM) ontogeny revealed the normal development of somatic embryos from globular to heart-shaped and dissection microscopy from torpedo-shaped to cotyledonary-stage embryos. The maximum germination percentage of 48% could be obtained on MS medium containing kinetin(1.0 mg $1^{-1}$) and the maximum survival percentage(79%) of plantlets after four weeks was found to be in the mixture of vermiculite, peatmoss, and soilrite(1:1:1).

• Plant Biotechnology Reports
• /
• 제4권4호
• /
• pp.269-280
• /
• 2010

Lilium ${\times}$ formolongi was genetically engineered by Agrobacterium-mediated transformation with the plasmid pCrtZW-N8idi-crtEBIY, which contains seven enzyme genes under the regulation of the CaMV 35S promoter. In the transformants, ketocarotenoids were detected in both calli and leaves, which showed a strong orange color. In transgenic calli, the total amount of carotenoids [133.3 ${\mu}g/g$ fresh weight (FW)] was 26.1-fold higher than in wild-type calli. The chlorophyll content and photosynthetic efficiency in transgenic orange plantlets were significantly lowered; however, after several months of subculture, they had turned into plantlets with green leaves that showed significant increases in chlorophyll and photosynthetic efficiency. The total carotenoid contents in leaves of transgenic orange and green plantlets were quantified at 102.9 and 135.2 ${\mu}g/g$ FW, respectively, corresponding to 5.6- and 7.4-fold increases over the levels in the wild-type. Ketocarotenoids such as echinenone, canthaxanthin, 3'-hydroxyechinenone, 3-hydroxyechinenone, and astaxanthin were detected in both transgenic calli and orange leaves. A significant change in the type and composition of ketocarotenoids was observed during the transition from orange transgenic plantlets to green plantlets. Although 3'-hydroxyechinenone, 3-hydroxyechinenone, astaxanthin, and adonirubin were absent, and echinenone and canthaxanthin were present at lower levels, interestingly, the upregulation of carotenoid biosynthesis led to an increase in the total carotenoid concentration (+31.4%) in leaves of the transgenic green plantlets.

• 한국초지조사료학회지
• /
• 제33권3호
• /
• pp.167-170
• /
• 2013

This study examined the growth performance and field evaluation of the dual herbicide-resistant transgenic creeping bentgrass plants. The effect of glyphosate treatment on the herbicide resistance of the transgenic creeping bentgrass plants was determined, and the non-transgenic control plant withered at the concentration $11{\mu}g/mL$ or higher whereas the transgenic creeping bentgrass plants survived the treatment at the concentration of $3,000{\mu}g/mL$, and the increase of the plant length was repressed as the glyphosate treatment concentration was increased. At field evaluation, glufosinate-ammonium and glyphosate were simultaneously treated to investigate the weed control effect. The results showed that more than 90% of the weeds withered four week after herbicide treatment, while the transgenic creeping bentgrass plants continued to grow normally. Therefore, the dual herbicide-resistant creeping bentgrass plants may be able to greatly contribute to the efficiency of weed control and to the economic feasibility of mowing in places such as golf courses.

• 한국발생생물학회지:발생과생식
• /
• 제6권2호
• /
• pp.83-88
• /
• 2002

월동기 90일 동안 transgenic coho salmon의 사료섭식량, 성장도 및 혈중 호르몬 수준을 파악하였다. 실험 종료시 실험구별 사료섭식량은 비교해 볼 때 tGH구가 Wild구보다 약4배 정도 높게 나타났지만, 사료효율의 경우 오히려 tGH구가 Wild구보다 1.1 배 낮은 것을 알 수 있었다. 한편 생존률은 tGH구가 Wild구보다 높게 나타났고, 성장은 tGH구가 길이에 있어 1.4배, 무게의 경우 약 3배 빠르게 나타났으며, 비만도 역시 tGH구가 Wild구보다 높게 나타났다. 실험구별 성장에 따른 호르몬 분석 결과, GH와 thyroid hormone의 농도는 tGH구가 Wild구에 비해 약 2배 정도 높은 값을 나타냈었다. 실험구별 형태적 특징을 관찰해 본 결과, tGH구들은 Wild구와 비교해 머리가 몸에 비해 상대적으로 크면서 꼬리지느러미의 가랑이 부분이 중심선으로 위, 아래가 확연히 분리되어 있지 않았다. 또한 몸통 부분은 길이에 비해 상대적으로 체고가 높게 나타났다.

• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2002년도 국제심포지엄
• /
• pp.101-101
• /
• 2002

GH-transgenic coho salmon (Oncorhynchus kitsutch) juveniles in tGH＊T$_3$and tGH*PTU were fed with the diets containing 1 ug/g fish of 3,5,3'-triiodo-L-thyronine (T$_3$) and 30 ug/g fish of 6-n-propyl-2- thiouracil (PTU), respectively, to assess the effect of these drugs on the change of physiological activity, growth and survival rate in comparison with normal transgenic (tGH＊C) and nontransgenic coho salmon (Wild) for 90 days. Although the daily food intakes of all transgenic (tGH)-groups were higher than Wild, the amount was reduced by exogenous PTU supply. The fred efficiencies of tGH-groups were lower than Wild, but the efficiency was reduced both by T$_3$and PTU. The survival rate of tGH-group was significantly higher than that of Wild, but there was no significant difference among tGH-groups. Although the growth of tGH-coho salmon was faster than Wild. the growth rate of transgenic salmon was increased by exogenous T$_3$, but was reduced by PTU Plasma TT$_4$levels of tGH-groups was approximately 2-fold higher relative to Wild, but there were no difference of plasma TT$_4$levels among tGH-groups. plasma TT$_3$level or tGH-coho salmon was increased by exogenous T$_3$administration, but was reduced by exogenous PTU. In addition, although plasma GH levels of all tGH-groups were higher than that of Wild, the GH level in plasma of transgenic coho salmon was increased by exogenous T$_3$and reduced by exogenous PTU. In the meantime, the transgenic fishes also displayed head, jaw and opercular abnormalities typical of the offsets of this gene construct in coho salmon, indicating that some imbalance in growth processes has been induced. However, the abnormalities of transgenic coho salmon was reduced following exogenous PTU administration.

• Journal of Microbiology and Biotechnology
• /
• 제23권12호
• /
• pp.1737-1746
• /
• 2013

IbMYB1, a transcription factor (TF) for R2R3-type MYB TFs, is a key regulator of anthocyanin biosynthesis during storage of sweet potatoes. Anthocyanins provide important antioxidants of nutritional value to humans, and also protect plants from oxidative stress. This study aimed to increase transgenic potatoes' (Solanum tuberosum cv. LongShu No.3) tolerance to environmental stress and enhance their nutritional value. Transgenic potato plants expressing IbMYB1 genes under the control of an oxidative stress-inducible peroxidase (SWPA2) promoter (referred to as SM plants) were successfully generated through Agrobacterium-mediated transformation. Two representative transgenic SM5 and SM12 lines were evaluated for enhanced tolerance to salinity, UV-B rays, and drought conditions. Following treatment of 100 mM NaCl, seedlings of SM5 and SM12 lines showed less root damage and more shoot growth than control lines expressing only an empty vector. Transgenic potato plants in pots treated with 400 mM NaCl showed high amounts of secondary metabolites, including phenols, anthocyanins, and flavonoids, compared with control plants. After treatment of 400 mM NaCl, transgenic potato plants also showed high DDPH radical scavenging activity and high PS II photochemical efficiency compared with the control line. Furthermore, following treatment of NaCl, UV-B, and drought stress, the expression levels of IbMYB1 and several structural genes in the flavonoid biosynthesis such as CHS, DFR, and ANS in transgenic plants were found to be correlated with plant phenotype. The results suggest that enhanced IbMYB1 expression affects secondary metabolism, which leads to improved tolerance ability in transgenic potatoes.

• 한국가축번식학회지
• /
• 제22권3호
• /
• pp.237-244
• /
• 1998

To investigate the fidelity of transgene transmission and expression, we produced transgenic mice carrying bovine $\beta$-casein/bovine grwoth hormone(bGH) fusion gene and examined transmission efficiency and expression level of the transgene in the founders and their progeny. The transgene was composed of 1.8 kb bovine $\beta$-casein promoter and 2.1 kb bGH gene. Ten transgenic mice were produced. Milk and mammary gland were collected from eight transgenic lines at 10-day lactation and a, pp.ied to Western and Northern blot analyses. The bGH expression was detected in four of them. The concentrations of bGH in milk were highly variable from 4$\mu\textrm{g}$/ml to 600$\mu\textrm{g}$/ml depending on the lines. The bGH mRNA level in mammary gland was closely correlated with the bGH concentration in milk in each transgenic line. These results indicated that bGH transgene expression was a, pp.opriately regulated in the mammary gland and secreted into milk in transgenic mice. By using two transgenic lines(#2, #7) secreting a considerable amoung of bGH into their milk, the inferitance and maintenance of transgenic phenotype were assessed in successive four generations. The mean transmission frequencies of transgene in lines #2 and #7 were 34% and 40%, respectively. The bGH concentration in milk were 80, 240, 120, 60$\mu\textrm{g}$/ml in each G0(generation 0), G2, G3, G4 generation of line #2 and 600, 1600, 860, 900$\mu\textrm{g}$/ml in each G1. G2, G3, G4 generation of line #7. These results demonstrated that bovine $\beta$-casein/bGH gene was stably transmitted from generation to generation in a Menelian fashion in trasgenic mice and consistenly expressed in their milk throughout the generations, although there was a little variation in the transmission frequency and expression level of the transgene between generations.