• 한국가축번식학회지
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• 제22권2호
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• pp.145-152
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• 1998

Many trials have been made to produce transgenic animals using sperm cells as a vector transferring foreign DNA into eggs, but reliable results are yet to be obtained (Brinster et al., 1989; Lavitrano et al., 1989; Bachiller et al., 1991; Sato et al., 1994). Recently, one of author(SO) demonstrated that mouse blastocysts derived from eggs fertilized by spermatozoa of male mice single injected with liposome-DNA complexes within the testis expressed thegene (Ogawa et al., 1995.) Here we report that a single injection of liposome-encapsulated DNAs into the testis of either male rats or mice resulted in successfully gene transfer to the postpartum progeny. The expression of mRNA derived from transgenes was also demonstrated in transgenic animals thus obtained. Further, the transmission of the exogenous gene to the descedants was confirmed in one line of transgenic rat up to F4 generation, indicating that the gene was stably incorporated into the germ line. Thus, direct single injection of foreign DNA into the testis provides a novel and convenient means to generate transgenic animals.

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

• Journal of Animal Science and Technology
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• 제47권4호
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• pp.547-554
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• 2005

본 연구는 메주에서 순수 분리된 메주곰팡이의 대표적 균종인 황국균(Aspergillus oryzae; AO)으로 만든 AO culture와 Salmonella 병원특이 유전자를 삽입한 형질전환 AO(TAO) culture가 산란계의 생산성, 계란 품질 및 장내 미생물 균총에 미치는 영향을 규명하고자 실시하였다. 39주령 산란계 Hy-line Brown 840수를 공시하여 대조구, AO culture 0.2%와 0.5%, TAO culture 0.2%와 0.5%, UV를 조사하여 단백질 분해효소를 감소시킨 mutant에 Salmonella 병원 특이 유전자를 삽입한 형질전환 AO(TMAO) culture 0.2%와 0.5% 첨가구들을 비교하였다. 각 첨가구는 6반복, 반복당 20수씩, 한 케이지 당 2수씩 배치하여 8주간 사양시험을 실시하였다. 사양시험 결과 모든 산란 생산성 및 계란 품질 관련 조사항목에서 처리간에 유의한(P<0.05) 차이가 있었다. TAO culture 0.2% 첨가구가 산란 생산성에 있어서 유의적으로 가장 높았으며, 난중은 모든 AO 첨가구들이 대조구에 비해 유의적으로 낮거나 낮아지는 경향을 나타내었다. 연파란율은 TMAO culture 0.5% 첨가구가 가장 낮았다. 사료섭취량과 사료요구율은 대조구와 모든 AO 첨가구들간에 유의적 차이가 나타나지 않았다. 난각 강도는 대조구 보다 모든 AO 첨가구들에서 유의적으로 높게 나타났으며, 난황 색도는 TMAO culture 0.5% 첨가구에서 가장 높았다. 난각 색도와 Haugh unit은 대조구와 모든 AO 첨가구들간에 유의적 차이가 나타나지 않았다. 장내 미생물 균총(Salmo- nella spp., E. coli. Lactobacilli spp.)에서는 유의적(P<0.05) 차이가 있었다. AO culture 첨가에 의해서 Lactobacilli spp.의 수는 증가되고, E. coli 및 Salmonella spp.의 수는 감소되었다. 특히 TAO와 TMAO culture 첨가구에서는 AO culture 첨가구보다 Salmonella spp. 및 E. coli 억제효과가 컸으며 첨가수준(0.5% vs 0.2%) 간에는 유의한 차이가 없었다. 결론적으로 TAO culture 0.2% 첨가는 산란 생산성 증가에 효과가 있었으며 TAO 및 TMAO culture 0.2% 첨가는 장내 E. coli 및 Salmonella spp.의 감소에 유의한 효과가 있었다.

• 한국동물생명공학회지
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• 제34권2호
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• pp.65-69
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• 2019

Since the development of the first genetically-modified mouse, transgenic animals have been utilized for a wide range of industrial applications as well as basic research. To date, these transgenic animals have been used in functional genomics studies, disease models, and therapeutic protein production. Recent advances in genome modification techniques such zinc finger nuclease (ZFN), transcription activator-like effector nucleases (TALEN), and clustered regularly interspaced short palindromic repeats (CRIPSR)-Cas9, have led to rapid advancement in the generation of genome-tailored livestock, as well as experimental animals; however, the development of genome-edited poultry has shown considerably slower progress compared to that seen in mammals. Here, we will focus primarily on the technical strategies for production of transgenic and gene-edited chickens, and their potential for future applications.

• Asian-Australasian Journal of Animal Sciences
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• 제21권6호
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• pp.801-806
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• 2008

A classical technique to study somitic cell fate is to employ the cross-transplantation of quail somites into a chick host. The densely stained nucleoli of the quail cells makes it possible to assess the fate of the donor quail cells in the chick host. Classical somite transplantation techniques have been hampered by the necessity of a small opening in the chick eggshell, difficulty in hatching the offspring and interspecies post-hatch graft rejection. With the advent of transgenic chicken technology, it is now possible to use embryos from transgenic chickens expressing reporter genes in somite cross-transplantation techniques to remove any possibility of interspecies graft rejection. This report describes using a surrogate eggshell system in conjunction with transgenic chick:chick somitic cell cross-transplantation to generate viable chimeric embryos and offspring. Greater than 40% of manipulated embryos survive past 10 days of incubation, and ~80% of embryos successfully cultured past 10 days of incubation hatched to produce viable offspring.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2006년도 제23차 정기총회 및 학술발표회
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• pp.74-76
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• 2006

닭의 간은 해독작용, 당의 저장, 혈장 단백질의 합성 등 주요 기능을 하는 것으로 알려져 있다. 본 연구는 간에서 성장 단계별로 발현량에 차이를 보이는 단백질들을 비교해 보았다. 2차원적 전기 영동에 의해 분리된 300개 이상의 단백질들이 확인되었으며 이 중 성장 단계별 특이적인 13개의 단백질은 MALDI-TOF MS에 의하여 분석이 되어졌다. 본 연구를 통하여 밝혀진 단백질들은 생화학적인 연구에 중요한 자료를 제공할 것으로 사료된다.

• Reproductive and Developmental Biology
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• 제33권2호
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• pp.107-111
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• 2009

The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{\times}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2004년도 제4회 발생공학 국제심포지움 및 학술대회
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• pp.45-45
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• 2004

• 한국가축번식학회지
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• 제22권3호
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• pp.237-244
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• 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.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.197-201
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• 2011

This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD45 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace ${\times}$ Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm, 75.4%), the fusion rate of the GalT/CDl6 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less then 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.