• 한국수정란이식학회지
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• 제27권1호
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• pp.9-14
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• 2012

This study was conducted to analyze the transgenic efficiency and sex ratio in ${\alpha}$-1,3-galactosyltransferase (GalT) knock-out (KO) transgenic pigs according to generation. GalT KO piglets were produced by artificial insemination or natural mating. The transgenic confirmation of GalT KO was evaluated by PCR amplification using specific primers. After electrophoresis, three types of bands were detected such as 2.3 kb single band (Wild), 2.3 and 3.6kb double bands (GalT KO -/+; heterozygote), and 3.6kb single band (GalT KO -/-; homozygote). Transgenic efficiency in F1 generation was 64.5% (23/35) of GalT KO (-/+). In F2 generation, GalT KO transgenic efficiency was 36.4% (21/57, Wild), 47.5% (28/57, GalT KO -/+), and 16.1% (8/57, GalT KO -/-), respectively. Interestingly, no homozygote piglets were born in 6 deliveries among total 11 deliveries, although they were pregnant between male (M) and female (F) $F_1$ heterozygote. In the 5 litters including at least one GalT KO -/- piglet, the transgenic efficiency was 13.3% (2/24, Wild), 51.3% (14/24, GalT KO -/+), and 35.3% (8/24, GalT KO -/-), respectively. The sex ratio of M and F was 40:60 in $F_1$ and 49:51 in $F_2$ generation, respectively. Based on these results, GalT KO transgenic pigs have had a reproductive ability with a normal range of transgenic efficiency and sex ratio.

• Asian-Australasian Journal of Animal Sciences
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• 제29권7호
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• pp.925-937
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• 2016

In the last few decades, transgenic animal technology has witnessed an increasingly wide application in animal breeding. Reproductive traits are economically important to the pig industry. It has been shown that the bone morphogenetic protein receptor type IB (BMPR1B) A746G polymorphism is responsible for the fertility in sheep. However, this causal mutation exits exclusively in sheep and goat. In this study, we attempted to create transgenic pigs by introducing this mutation with the aim to improve reproductive traits in pigs. We successfully constructed a vector containing porcine BMPR1B coding sequence (CDS) with the mutant G allele of A746G mutation. In total, we obtained 24 cloned male piglets using handmade cloning (HMC) technique, and 12 individuals survived till maturation. A set of polymerase chain reactions indicated that 11 of 12 matured boars were transgene-positive individuals, and that the transgenic vector was most likely disrupted during cloning. Of 11 positive pigs, one (No. 11) lost a part of the terminator region but had the intact promoter and the CDS regions. cDNA sequencing showed that the introduced allele (746G) was expressed in multiple tissues of transgene-positive offspring of No.11. Western blot analysis revealed that BMPR1B protein expression in multiple tissues of transgene-positive $F_1$ piglets was 0.5 to 2-fold higher than that in the transgene-negative siblings. The No. 11 boar showed normal litter size performance as normal pigs from the same breed. Transgene-positive $F_1$ boars produced by No. 11 had higher semen volume, sperm concentration and total sperm per ejaculate than the negative siblings, although the differences did not reached statistical significance. Transgene-positive $F_1$ sows had similar litter size performance to the negative siblings, and more data are needed to adequately assess the litter size performance. In conclusion, we obtained 24 cloned transgenic pigs with the modified porcine BMPR1B CDS using HMC. cDNA sequencing and western blot indicated that the exogenous BMPR1B CDS was successfully expressed in host pigs. The transgenic pigs showed normal litter size performance. However, no significant differences in litter size were found between transgene-positive and negative sows. Our study provides new insight into producing cloned transgenic livestock related to reproductive traits.

• 한국임상수의학회지
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• 제27권4호
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• pp.330-335
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• 2010

이 연구의 목적은 돼지에 있어서 zoletil/midazolam (TZM)과 zoletil/xylazine (TZX)의 2가지 병용마취를 사용하였을 때의 마취효과와 심혈관계 및 호흡에 미치는 영향을 비교하기 위하여 실시하였다. 총 8 마리의 Landrace $\times$ Yorkshire 교잡종 돼지 ($25.3{\pm}3.3\;kg$)를 사용하였으며, 각 군마다 4 마리씩 2개 군으로 실험을 실시하였다. 첫 번째 군은 xylazine과 tiletamine/zolazepam을 2 mg/kg 용량으로 근육 내 주사하였고(TZX군), 2번째 군은 midazolam 0.5 mg/kg의 용량을 정맥 내 주사 하고 tiletamine/zolazepam 2 mg/kg 용량으로 근육 내 주사하였다(TZM군). 마취시간에 대한 평가로 induction time, anesthesia time과 standing time을 각 돼지마다 측정하였으며, 마취효과에 대한 평가로 진정, 진통, 근 이완, 자세 그리고 청각반응을 점수화 하여 매 15분마다 측정하였다. 심폐기능에 대한 평가로 심박동수와 동맥혈압, 호흡수, 직장체온을 마취 전, 마취직후, 마취 후 5분, 15분, 30분, 45분 및 60분에 각각 측정하였고, 동맥혈 가스 분석을 동일시간대에 실시하였다. 실험 결과 모든 돼지에서의 마취는 성공적이었다. 2가지 병용마취 모두 부드러운 마취 유도와 적절한 운동억제 효과를 보였으나 마취효과점수에서는 TZM군이 TZX군보다 우수하였으며, TZM군이 TZX군 보다 심폐기능 및 체온에 미치는 영향이 적으며 안정적인 것을 확인하였다. 결론적으로, TZM군은 TZX군에 비하여 더욱 양호한 마취효과를 보였으며, 심폐기능에 미치는 영향은 보다 적게 나타내었다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권10호
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• pp.1478-1484
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• 2007

To determine the origin and genetic diversity of Red Chittagong (RC) cattle in Bangladesh, we analyzed mitochondrial DNA displacement loop (D-loop) sequences of 48 samples along with 22 previously published sequences from Bos indicus and Bos taurus breeds. Twenty five haplotypes were identified in RC cattle that were defined by 44 polymorphic sites and nucleotide diversity was $0.0055{\pm}0.0026$. The estimated sequence divergence times between RC and other zebu cattle breeds studied ranged between 22,700-26,900 years before present (YBP) which, it is suggested, predate domestication of RC cattle. Furthermore, it is assumed that introgressions have occurred in this breed mainly from Indian zebu breeds in the recent millennia. The phylogenetic studies showed RC cattle clustered with Bos indicus lineage with two distinct haplogroups representing high genetic variability of this breed. These findings can be used for designing proper breeding and conservation strategies for RC cattle in Bangladesh.

• 한국수정란이식학회지
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• 제30권4호
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• pp.305-313
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• 2015

Xenotransplantation of pig islet regarded as a good alternative to allotransplantation. However, cellular death mediated by hypoxia-reoxygenation injury after transplantation disturb success of this technique. In the present study, we produce transgenic pig expressing human heme oxygenase 1 (HO1) genes to overcome cellular death for improving efficiency of islet xenotransplantation. Particularly, Korean miniature pig breed, Micro-Pig, was used in the present study. Somatic cell nuclear transfer (SCNT) technique was used to produce the HO1 transgenic pig. Six alive transgenic piglets were produced and all the transgenic pigs were founded to have transgene in their genomic DNA and the gene was expressed in all tested organs. Also, in vitro cultured fibroblasts derived from the HO1 transgenic pig showed low reactive oxygen species level, improved cell viability and reduced apoptosis level.

• 한국임상수의학회지
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• 제28권3호
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• pp.291-296
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• 2011

돼지에서 medetomidine-ketamine (MK) 합제에 대한 마취효과와 이 합제에 대한 atipamezole (MKA) 과 yohimbine (MKY)의 길항효과를 비교하였다. 24 마리 Landrace - Yorkshire 혼혈 종 돼지를 사용하였다. Medetomidineketamine 는 한 주사기로 근육주사 하였고 atipamezole 과 yohimbine 은 마취 후 20 분에 정맥 주사 하였다. 평균마취시간, 평균흉와시간, 평균기립시간 및 평균보행시간은 MKA와 MKY군에서 MK군보다 유의적으로 짧았다. 그러나 MKA군과 MKY군간의 유의적인 차이는 없었다. 평균혈압은 MKA와 MKY군에서 MK군보다 유의적으로 낮았다. 결론적으로 Medetomidine-ketamine 에 의한 마취 및 혈압 상승 효과는 atipamezole과 yohimbine 에 의해 안전하고 빠르게 길항되었다. 따라서 atipamezole과 yohimbine은 돼지에서 Medetomidine-ketamine 마취를 길항하는 데 안전하고 효과적으로 사용될 수 있다.

• 한국수정란이식학회지
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• 제32권3호
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• pp.227-233
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• 2017

Pig has been known to be one of the most feasible animals as a bioreactor to produce pharmaceuticals in milk and as a mediator in xenotransplantation research. Previously, we generated transgenic pigs for both purposes, which were expressing Factor 8, vWF, hTPA, and hEPO in milk, along with expression of MCP at GalT gene locus ($GalT^{-MCP/-MCP}$) as well as expressing MCP at GalT gene loci with CD73 expression ($GalT^{-MCP/+}/CD73$). In this study, we performed comparative analyses of sperm parameters between wild type male (WT) pig and those transgenic males to examine the effects of transgenes integrated into the pigs on motility, morphology, viability, and acrosome integrity of the spermatozoa. Our results showed that the rates of actively motile spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 85.0%, 83.3%, 82.5%, 83.3%, 82.5%, 77.5%, and 78.7%, respectively. Whereas, the rates of morphologically normal spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 90.0%, 80.0%, 80.0%, 83.3%, 85.0%, 91.8%, and 80.8%, respectively. In addition, the viability in spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 93.9%, 82.4%, 89.9%, 83.9%, 87.4%, 92.8%, and 83.6%, respectively. The rates of spermatozoa with normal acrosome integrity in WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 98.1%, 98.6%, 98.6%, 98.7%, 98.1%, 99.5%, and 95.1%, respectively. There were no significant differences in motility, morphology, viability, and acrosome integrity of the spermatozoa among WT, Factor 8, vWF, hTPA, and hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs. These mean that neither random integration nor targeted integration of the transgene into chromosome of pig effect on characteristics of spermatozoa. Ultimately, the transgenic male pigs subjected in this study could apply to propagate their progenies for production of human therapeutic proteins and advancing the xenotransplantation research.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.233-233
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• 2004

PURPOSE: Gene expression and apoptosis in testicular germ cells has been demonstrated in many transgenic animals. However, little is known about the transgenic pig and rates of apoptosis during spermatogenesis. METHODS : Morphological and biochemical features of apoptosis reported in other species were used to confirm that the TdT-mediated dUTP Nick end labeling (TUNEL) assay is an acceptable mothos for idendtification and quantification of apoptotic transgenic germ cells in histological tissue section from transgenic pig testis. (omitted)

• 대한약학회:학술대회논문집
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• 대한약학회 2005년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.106-112
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• 2005

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2004년도 제4회 발생공학 국제심포지움 및 학술대회
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• pp.18-19
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• 2004