• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2000년도 추계학술대회 및 정기총회
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• pp.46-54
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• 2000

Transgenic rabbits were produced by DNA microinjection using growth hormone receptor (GHR) and IGF-1 receptor (IGF-1R) genes. Overall efficiencies for production of transgenic rabbits were 3.2% and 3.1% in GHR and IGF-1R genes, respectively. Founder rabbits transmitted transgenes to their progenies through medelian fashion. Growth rate in GHR and ICF-1R transgenic rabbits was faster than non-transgenic rabbits. Transgenic rabbits grew larger (25% and 15% increase in body weight of GHR and IGF-1R transgenic rabbits, respectively) than non-transgenic rabbits and organ weight of transgenic rabbits increased, suggesting that GHR and IGF-1 genes affects growth rates in transgenic rabbits.

• 한국가축번식학회지
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• 제27권1호
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• pp.1-7
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• 2003

MT-GHR(Growth hormone receptor)와 MT-IGF-IR(IGF-1 receptor)유전자를 구축하고 micromani-pulator를 이용하여 토끼 수정란에 유전자를 주입하여 형질전환토끼를 생산하였다. 본 연구에서의 형질전환토끼의 생산효율은 약 3%를 나타내었고 Growth Hormone receptor(GHR)를 가진 형질전환 토끼와 IGF-1 receptor(IGF-lR)를 가진 형질전환토끼를 10마리 이상씩 생산하였다. 또한 정상 토끼와 교배시켜 F$_1$ 새끼를 얻어 유전자가 다음세대에도 전달되는 것을 확인하였다. GHR 이나 IGF-1R 형질전환토끼의 성장률은 정상토끼보다는 약15~25% 정도 빠른 경향을 나타냈고 특히 GHR 형질전환토끼의 성장률이 더 높은 것으로 드러나 GHR 및 IGF-lR유전자가 형질전환토끼에서 성장에 영향을 주었다는 것을 확인할 수 있었다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.95-95
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• 2002

Transgenic rabbits were produced by micoinjection of DNA containing metallothionein promoter ligated to growth hormone receptor (GHR) and IGF-l receptor (IGF-lR) genes. Founder transgenic rabbits transmitted transgenes into pups with Medelian ratio. The mRNA expression of transgenes using Northern blotting with probes of IGF-IR and GHR genes was checked in liver of transgenic rabbits. Transgenic rabbits with IGF-IR and GHR genes more expressed mRNA than control non-transgneic rabbits. (omitted)

• Asian-Australasian Journal of Animal Sciences
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• 제20권4호
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• pp.482-486
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• 2007

Basic objective of this research was to compare the milk yield and composition of New Zealand White transgenic rabbit females expressing recombinant human factor VIII (hFVIII) in mammary gland during lactation with that of non-transgenic rabbit females of the same age during 30 days of lactation. Transgenic founders were generated by the microinjection of foreign DNA (mWAP-hFVIII gene construct) into the egg. F1, F2 and F3 generations of transgenic rabbits were obtained after mating of transgenic founder rabbits with non-transgenic rabbits. The amount of milk rejected was measured by weight-suckle-weight method at $10^{th}$, $20^{th}$and $30^{th}$ day of lactation. Quality of milk (content of fat, protein, lactose, dry ash, and some minerals) from transgenic and non-transgenic rabbit was also determined. Comparison of milk yield, determined by weight-suckle-weight method, showed significantly higher (p<0.05) milk production at day 20 of first lactation in non-transgenic females, but on the same day of second lactation higher milk yield was measured in transgenic ones. Significantly higher (p<0.05) content of milk fat and protein was determined in transgenic milk whilst higher content of lactose was found in non-transgenic milk. The content of minerals (calcium, phosphorus, magnesium and sodium) did not differ in both experimental and control groups. Our results showed that milk yield and composition of transgenic rabbit females (mammary specific transgenic over-expression of hFVIII) over several generations is only slightly and transiently different from milk yield of non-transgenic females, which had no significant consequence on the litter size and viability.

• 생약학회지
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• 제31권2호
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• pp.149-156
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• 2000

Atherosclerosis is emerging as one of the major causes of death in Korea as well as Western societies. In the present study; hypocholesterolemic and antiatherogenic effects of the ethanol extract of Allium victorialis Makino was investigated using the conventional rabbit and the cholesteryl ester transfer protein (CETP)-transgenic mouse model. Hypercholesterolemia was induced by feeding high cholesterol diet to the animals for 30 days and they were then fed with high cholesterol diet containing 0.5% of the A. victorialis extract for additional 30 (or 40) days. In the experiment using rabbits, treatment with the A. victorialis extract significantly decreased plasma total cholesterol, low density lipoprotein (LDL)-cholesterol, triglyceride levels and lipid peroxidation compared to those in the control group. Total cholesterol contents in the liver and the heart were also significantly decreased. Lipid staining of the aorta isolated from the rabbits showed that treatment with the A. victorialis extract decreased formation of atheromatous plaques on the intima of the aorta. In the experiment employing CETP transgenic mouse model, treatment with the A. victorialis extract decreased the levels of plasma total cholesterol and the tissue triglyceride levels in the heart. These results demonstrated that the ethanol extract of A. victorialis lowered serum cholesterol levels, tissue lipid contents and accumulation of cholesterol in the artery.

• 한국임상수의학회지
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• 제25권3호
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• pp.165-169
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• 2008

Renal ischemia-reperfusion (I/R) injury is great clinical important because viability of the organ depends on the tolerance to ischemia-reperfusion injury, an inevitable processing during surgery. The purpose of this study was to investigate the effects of premedicated ascorbic acid alone in I/R injury model induced by cross-clamping of renal vessels. In the rabbit models, 2-4 kg New Zealand white rabbits were subjected to 30 minutes of warm unilateral renal ischemia followed by removal of contralateral kidney and then divided into five groups, control (2) arid treatment groups (3). In control group 1, the rabbits only received right nephrectomy. In control group 2, the rabbits received I/R on left kidney after the right nephrectomy. In treatment group 1, the rabbits received ascorbic acid 50 mg/kg IV before the operation. In treatment group 2, the rabbits received ascorbic acid 100 mg/kg IV before the operation. In treatment group 3, the rabbits received ascorbic acid 200 mg/kg IV before the operation. Blood samples were collected from these rabbits for measurement of kidney function tests at the 0, 1 st, 3rd and 7th day and antioxidant enzyme( SOD, GSHPx, CAT) at 24 hours. Kidney function tests (serum creatinine and BUN) showed a significant difference between group 2 and group 4, 5. Activity of antioxidant enzymes in plasma were significant decrease in group 4, 5 compare to group 2. The result of this study suggested that the exogenous ascorbic acid had a role of attenuation of renal I/R injury in rabbit model.

• 한국수정란이식학회지
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• 제13권2호
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• pp.97-106
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• 1998

The pronuclear injection of metallothionein-human growth hormone (MT-hGH) gene into rabbit zygotes was performed to establish in vitro developmental system and to detect the presence of the injected gene by nested PCR. Mature female New Zealand White rabbits were superovulated by eGG and hCG treatments. The rabbits were mated and the zygotes were collected from the oviducts 18-22 h after hCG injection by flushing with D-PBS. Two to three picoliters of MT-hGH gene was microinjected into male pronuclei. The foreign gene-injected zygotes were cultured in TCM-199 or RD mediurn containing 10% FCS with a monolayer of rabbit oviductal epithelial cefls in a 5% $CO_2$ incubator. The presence of injected DNA in rabbit embryos or blastomeres at different developmental stages .vas detected by a nested PCR analysis. The results are summarized as follows ; 1.The developmental rate of the MT-hGH gene-injected zygotes to blastocyst was significantly higher in TCM-199 medium (68.1%) than in RD medium (42.9%). 2.The gene injection into pronuclei at 18 or 22 hours post hCG treatment during pronuclear stage did not much affect on the in vitro development of the rabbit embryos. 3.The rate of gene-positive embryos detected by the nested PCR analysis was significantly decreased when they developed to blastocysts. The results indicate that the screening of transgene in rabbit embryos by nested PCR analysis could be a prornisible method for the preselection of transgenic embryos. Furthermore, the preselection of transgenic embryos would greatly reduce hoth the cost and effort of production of transgenic animals.

• 한국수정란이식학회지
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• 제15권2호
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• pp.167-173
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• 2000

The principal objective of this study was to clone transgenic embryos in order to improve the efficiency of transgenic animal production by the combination of microinjection and nuclear transplantation techniques. Mature female New Zealand White rabbits were superovulated by eCG and hCG treatments, fllowed by natural mating. Zygotes were collected from the oviducts at 18∼22 h after hCG injection by flushing with D-PBS containing 5% fetal calf serum(FCS). Two to three picoliters of green fluorescent protein(GFP) gene wa microinjected into male pronucleus. The foreign gene-injected zygotes were cultured in TCM-199 or RD medium containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% CO2 incubator. The morulae expressing GFP gene were selected and their blastomeres were separated for the use of nuclear donor. Following nuclear transplantation of fluorescence-positive morula stage blastomeres, 13 (21.3%) out of 61 fused oocytes developed to blastocyst stage and all of the cloned blastocysts expressed GFP. The results indicate that the screening of transgene in rabbit embryos by GFP detection could be a promisible method for the preselection of transgenic embryos. Also the cloning of preselected transgenic embryos by nuclear transplantatin could be efficiently applied to the multiple production of transgenic animals.

• BMB Reports
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• 제35권2호
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• pp.172-177
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• 2002

We previously reported that cholesteryl ester transfer protein (CETP) inhibitory peptides (designated $P_{28}$ and $P_{10})$ have anti-atherogenic effects in hypercholesterolemic rabbits (Biochim. Biophys. Acta (1998) 1391, 133-144). To further investigate those effects, we studied rabbit plasma that was collected after 30 h of a $P_{28}$ or $P_{10}$ injection. We found that there is a strong correlation between the in vivo CETP inhibition effects and alterations of lipoprotein particle size distribution in rabbit plasma, as determined on an agarose gel electrophoresis and gel filtration column chromatography. In vivo effects of the peptide were observed again in C57BL/6 mice that expressed simian CETP. The $P_{28}$ or $P_{10}$ peptide ($7\;{\mu}g/g$ of body weight) that was dissolved in saline was injected subcutaneously into the mice. The $P_{28}$ injection caused the partial inhibition of plasma CETP activity up to 50%, decreasing the total plasma cholesterol concentration by 30%, and increasing the ratio of HD/total-cholesterol concentration by 150% in the CETP-transgenic (tg) mice. The CETP inhibition by the $P_{28}$ or $P_{10}$ made alterations that modulated the size re-distribution of the lipoproteins in the blood stream. Particle size of the very low (VLDL) and low density lipoproteins (LDL) from the peptide-injected group was highly decreased compared to the saline-injected group (determined on the gel filtration column chromatography). In contrast, The HDL particle size of the $P_{28}$-injected group increased compared to the control group (saline-injected). The expression level of the CETP mRNA of the $P_{28}$-injected CETP-tg mouse appeared lower than the saline-injected CETP-tg mouse. These results suggest that the injection of the CETP inhibitory peptide could affect the CETP expression level in the liver by influencing lipoprotein metabolism.

• Journal of Microbiology and Biotechnology
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• 제18권1호
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• pp.153-159
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• 2008

The limitations in current technology for generating transgenic animals, such as the time and the expense, hampered its extensive use in recombinant protein production for therapeutic purpose. In this report, we present a simple and less expensive alternative by directly infusing a recombinant adenovirus vector carrying human lactoferrin cDNA into rabbit mammary glands. The milk serum was collected from the infected mammary gland 48 h post-infection and subjected to a 10% SDS-PAGE and Western blotting. An 80-kDa protein was visualized after viral vector infection. With this method, we obtained a high level of expressed human lactoferrin of up to 2.3mg/ml in the milk. Taken together, the method is useful for the transient high-level expression recombinant proteins, and the approach established here is probably one of the most economical and efficient ways for large-scale production of recombinant proteins of biopharmaceutical interest.