• 제목/요약/키워드: Cloned pigs

검색결과 223건 처리시간 0.024초

사람 H-Transferase 유전자 과발현 형질전환 체세포주 확립 및 검증 (Production and Characterization of Porcine Cell Lines Overexpressing Human H-Transferase)

  • 이상미;박효영;김혜민;문승주;강만종
    • Reproductive and Developmental Biology
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    • 제31권3호
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    • pp.187-191
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    • 2007
  • 본 연구는 사람 H-transferase가 과발현하는 돼지 체세포주를 개발하는데 있다. 돼지 세포에 사람 H-transferase 유전자를 발현시키는 것은 이종간 장기 이식에 있어서 초급성 거부 반응을 방지하기 위한 한 가지 방법이다. 본 연구에서는 과발현 벡터를 구축하기 위하여 사람 H-transferase을 HepG2 세포로부터 동정하였으며, 이 유전자를 CMV promoter를 이용하여 발현할 수 있도록 포유동물 발현 벡터인 pRc/CMV벡터에 삽입하였다. 또한, 돼지 산자의 귀 세포를 이용하여 체세포를 수립한 후 jetPEI DNA transfection reagent를 이용하여 벡터를 도입하였고, $300{\mu}g/ml$의 G418로 12일간 선별하였다. PCR을 이용하여 선별된 colony들을 분석한 결과, 벡터가 도입되었음을 확인하였고, RT-PCR을 이용하여 사람 H-transferase mRNA가 발현하는 것을 확인하였다. 본 연구에서 확립된 세포주는 사람 H-transferase가 과발현하는 형질 전환돼지의 생산에 이용될 수 있을 것으로 생각된다.

Molecular Cloning, Segmental Distribution and Ontogenetic Regulation of Cationic Amino Acid Transporter 2 in Pigs

  • Zou, Shi-geng;Zhi, Ai-min;Zhou, Xiang-yan;Zuo, Jian-jun;Zhang, Yan;Huang, Zhi-yi;Xu, Ping-Wen;Feng, Ding-yuan
    • Asian-Australasian Journal of Animal Sciences
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    • 제22권5호
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    • pp.712-720
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    • 2009
  • The goal of this study was to elucidate the expression and segmental distribution of the glomerular cationic amino acid metabolism transporter-2 (CAT-2) and thus to improve our understanding of porcine cationic amino acid transporters and amino acid absorption. Porcine CAT-2 was cloned, sequenced and characterized. The predicted amino acid sequence of porcine CAT-2 shared 86.1% and 92.1% identity with human and mouse CAT-2A, respectively. The tissue distribution patterns and ontogenic changes of CAT-2 mRNAs were determined by real-time Q-PCR. The results showed that porcine CAT-2 was highly expressed in the heart and intestinal tract (duodenum, ileum and jejunum). In addition, the mRNA of CAT-2 was found in liver, lung, kidney, brain and muscle. Within the intestinal tract, CAT-2 mRNA was most abundant in the ileum and rarely expressed in the duodenum. In the duodenum, the levels of CAT-2 mRNA reached their peak on day 7 (p<0.05) while in the jejunum, levels were low on day 1 and 7 and increased rapidly after day 26 before peaking on days 30 and 60 (p<0.05). The levels then dramatically decreased by day 90 (p<0.05). In the ileum, levels achieved their maximum on day 30 and then decreased significantly on day 60 (p<0.05).

동물원에서 사육중인 Japanese Macaque (Macaca fuscata)에서 Zoletil 마취에 관한 연구 (Anesthetic Effects of Zoletil on Japanese Macaque (Macaca fuscata) Reared in Zoological Garden)

  • 이재일;이수진;홍성혁;신남식;김덕환;박창식;전무형;조종민;김영석;김명철
    • 대한수의학회지
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    • 제43권2호
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    • pp.301-305
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    • 2003
  • To evaluate the effects of Zoletil (tiletamine- zolazepam) that have been widely used for the chemical restraint and anesthesia of primates, on physiologic alteration, blood gas analysis and anesthetic duration in the Japanese macaque(Macaca fuscata), this study was carried out. Zoletil was administered by intramuscular injection. Evaluation of temperature, heart rate, respiration rate and blood gas analysis were performed before administration and at 1, 10, 20, 30, 40, 50 and 60 min after administration, and induction and maintenance time was recorded. There was no significant difference in heart rate, $PCO_2$, $PO_2$ after Zoletil administration rut temperature, respiraticn rate, pH were significant difference compared with these of Mere administration. The induction time was $2.5{\pm}1.0min$ and maintenance time was $86{\pm}23.2min$. It was considered that Zoletil could be usefully used for the sedation and immobilization of Japanese macaque reared in zoological garden.

Associations of Polymorphisms in the Mx1 Gene with Immunity Traits in Large WhitexMeishan F2 Offspring

  • Li, X.L.;He, W.L.;Deng, C.Y.;Xiong, Y.Z.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권11호
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    • pp.1651-1654
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    • 2007
  • The mouse myxovirus resistance protein 1 (Mx1) is known to be sufficient to confer resistance to influenza viruses, and the gene encoding Mx1 is, therefore, an interesting candidate gene for disease resistance in farm animals. The porcine Mx1 gene has already been identified and characterized based on its homology with mouse Mx1; the full-length coding region of the pig Mx1 gene spans 2,545 bp (M65087) and is organized into 17 exons compared with the human ortholog mRNA. In this study, the exons 9, 10 and 11 and introns 6 and 9 of the porcine Mx1 gene were cloned and sequenced. Two SNPs were identified in exons 9, 10 and 11 but none of the SNPs led to an amino acid exchange, and the other eleven variants were detected in introns 6 and 9, respectively. Differences in allele frequency between Meishan and other pig breeds were observed within intron 6, of which an $A{\rightarrow}G$ substitution at position 371 was detected as an SnaBI PCR-RFLP. The association analysis using the Large White${\times}$Meishan $F_2$ offspring suggested that the Mx1 genotype was associated with variation in several immunity traits that are of interest in pig breeding. However, further investigations in more populations are needed to confirm the above result.

Bacterial Diversity at Different Sites of the Digestive Tract of Weaned Piglets Fed Liquid Diets

  • Hong, Tran Thi Thu;Passoth, Volkmar;Lindberg, Jan Erik
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권6호
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    • pp.834-843
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    • 2011
  • Bacterial diversity was studied using PCR-DGGE, cloning and sequencing. DNA was isolated from digesta samples from stomach, ileum and colon of 28 weaned piglets (Large White${\times}$Mong Cai) fed dry control feed, naturally fermented liquid feed (FE) and a liquid diet with inclusion of rice distiller's residue feed. General bacterial diversity was described using DGGE analysis of the V3 region of 16S rDNA. The microbial populations in the stomach and the ileum were considerably influenced by the diet, while only marginal effects were observed in the colon. There was a large variation of the microbial flora in the stomach between individuals fed non-fermented diets. In contrast, animals fed diet FE had a more uniform microbial flora in the stomach and the ileum compared to the other diets. In total 47 bands from the DGGE profiles were cloned. In stomach, most frequently lactic acid bacteria were found. Feeding diet FE resulted in the occurrence of Pediococcus species in stomach and ileum. In pigs fed the other diets, Lactobacillus gallinarum, Lactobacillus johnsonii and Lactobacillus fermentum were found in stomach and ileum. Most of the sequences of bands isolated from colon samples and several from ileum matched to unknown bacteria, which often grouped within Prevotellaceae, Enterobacteriaceae, Bacteroidaceae and Erysipelotrichaceae. This study demonstrates that fermented liquid feed affects bacterial diversity and the specific microflora in stomach and ileum, which provides a potential to modulate the gut microflora with dietary means to increase the abundance of beneficial bacteria and improve piglets' health.

한우 섬유아세포의 성 판별 및 세포주기 유도 분석 (Sexing and Cell Cycle Induction Hanwoo Fetal Fibroblast Cells)

  • 김현주;강회성;최화식;이성호;박창식;진동일
    • 한국가축번식학회지
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    • 제27권1호
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    • pp.53-59
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    • 2003
  • 본 연구에서는 한우 태아의 시기별로 35일령, 50일령, 70일령 및 90일령의 fetal fibroblast cell line을 생산하였고, bovine-specific primer와 Y chromosome-specific primer를 이용하여 PCR에 의해 성을 판별하여 각각 암수 2 line의 한우 fetal fibroblast cell line을 확립하였다. 이들 cell line을 계대배양하여 passage number가 10 이상에서 염색체 분석을 실시하였는데 모두에서 80%이상의 세포가 60개의 정상 염색체수의 나타내어 계대배양이 karyotype에 영향을 미치지 않는 것으로 나타났다. Serum starvation과 confluent 배양 방법을 이용하여 Go 상태로 유도되었는지 확인하기 위해 PCNA antibody를 이용하여 Western blotting 분석을 실시하였는데 PCNA 발현이 현저히 감소되는 것을 확인할 수 있었고, 다시 정상 medium으로 환원시켰을 때 세포분열이 재개되어 Go상태로 유도되었음을 확인할 수 있었다. 또한 serum stravation 방법이 conflent한 배양방법보다 PCNA 발현양이 적은 것으로 나타나 좀더 효율적인 Go 상태 세포 주기 조절방법으로 판명되었다.

우유로부터 Angiogenin의 정제 (Purification of Angiogenin from Bovine Milk)

  • 남명수;배형철;박창식
    • Journal of Animal Science and Technology
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    • 제46권1호
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    • pp.77-82
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    • 2004
  • 본 연구는 우유의 angiogenin(ANG) 생리활성기능을 조사하기 위한 전단계로 ANG 정제 방법을 확립하기 위하여 실시하였다. 우유로부터 ANG을 CM-Sepharose ion exchange chromatography, HPLC, Gel-filtration의 3단계 방법을 이용하여 정제하였다. 1단계인 CM-Sepharose ion exchange chromatography를 수행 한 결과 0.6 M NaCl 농도의 36번 분획에서 ANG의 밴드를 볼 수 있었고 2단계인 Mono-S 컬럼을 이용하여 HPLC를 수행한 결과에서는 1.0 M에서 하나의 peak가 분리되었고 전기영동 결과는 LF와 ANG 두 가지 성분이 함유되어 있음을 확인하였다. 마지막 단계인 Gel-filtration을 수행한 결과 ANG이 단일 밴드로 분리되었다. 우유 10 $\ell$로부터 약 80 ${\mu}\ell$의 ANG를 얻었고 정제된 ANG을 N-말단 아미노산 배열을 분석한 결과 AQDDYRYIHFLTQHY로 밝혀졌다.

젖소 족피부염(足皮膚炎)에 대한 오존연고의 치료(治療) 효과(效果) (The Efficacy of Ozone Ointment Therapy on Pododermatitis of Dairy Cows)

  • 이수진;전무형;조성환;김덕환;박창식;김명철
    • 한국임상수의학회지
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    • 제23권3호
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    • pp.279-285
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    • 2006
  • Thirty dairy cows with pododermatitis were selected and treatment effect of ozone ointment for bovine pododermatitis was investigated. In addition, bactericidal effect of ozone ointment on etiological agent of bovine pododermatitis was evaluated. The pathohistological examination for the pododermatitis, according to application with ozone ointment was investigated. Thirty dairy cows were divided two groups: control group(vaseline group: 15 cows), treatment group(ozone ointment group: 15 cows). Various parameters were evaluated in terms of the lameness score, swelling score, lesion score, WBC, neutrophil, pathohistological finding, and antimicrobial action. As compared with vaseline group, ozone ointment group revealed significant decrease of lameness(p<0.05), swelling(p<0.01) and lesion score(p<0.05) were shown in hoof lesions on 14 days after application. In hematological findings, WBC count revealed slightly high values within normal range before treatment, however, this was 1Corresponding author improved on 14 days after application of ozone ointment. The number of neutrophils was slightly higher than that of normal, however, this was improved on 14 days after application of ozone ointment. In pathohistological findings, normal dermal tissue was found in tissues with pododermatitis on 14 days after application of ozone ointment. In antimicobial action, marked decrease rate of bacteria was observed in feet of all cases treated with ozone ointment. The decreasing rate of bacteria in anaerobic culture was higher than that in aerobic culture.

젖소 족피부염(足皮膚炎)에 대한 오존수의 치료(治療) 효과(效果) (The Efficacy of Ozonated Water Therapy on Pododermatitis of Dairy Cows)

  • 이수진;조성환;전무형;김덕환;박창식;한홍율;김명철
    • 한국임상수의학회지
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    • 제23권3호
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    • pp.272-278
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    • 2006
  • This study was carried out to determine therapeutic the effect of ozonated water therapy on bovine pododermatitis. In addition, bactericidal effect of ozonated water on etiological agent of bovine pododermatitis was examined. The pathohistological examination for the pododermatitis, according to application with ozonated water and ozone ointment was investigated. Thirty healthy cattle were divided two groups(each of 15) : control group(povidone group), treatment group(ozone solution group). Various parameters were evaluated in terms of the lameness score, swelling score, lesion score, WBC count, neutrophil count, pathohistological finding, and antimicrobial action. The decrease of lameness and lesion score were shown in hoof lesions on 14 days after application of ozonated water. Significant decrease of swelling was shown in hoof lesions on 14 days 1Corresponding author after application of ozonated water(p<0.01). In hematological findings, WBC count revealed values within normal range. The number of neutrophils was slightly higher than that of normal, however, this was improved on 14 days after application of ozonated water. In pathohistological findings, recovery was rapid macroscopically and microscopically in the treatment with ozonated water on the hoof lesions and ozonated water was effective. In antimicobial action, bactericidal effect was observed in treatment with ozonated water on the hoof lesions and ozonated water was effective.

Antioxidant Effects of Ascorbic Acid on Renal-Ischemia Reperfusion Injury in Rabbit Model

  • Kim, Jong-Man;Lee, Jae-Yeon;Kim, Duck-Hwan;Jeong, Seong-Mok;Park, Chang-Sik;Kim, Myung-Cheol
    • 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.