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섬유소 분해효소 유전자 도입 형질전환 돼지 생산

  • 이연근;박진기;민관식;성환후;임기순;양병철;김진회;류재웅;장원경
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.58-58
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    • 2001
  • 본 연구는 섬유소 분해효소 유전자 Cel D(Cellulase Digestion)가 도입된 형질전환 돼지 생산을 통하여 섬유소 함유 사료의 이용 효율을 증대시키고, 나아가 장기이식동물 및 고가의 의료용 단백질 생산가축을 개발하기 위한 원천기술 확보에 있다. 섬유소분해 유전자의 크로닝 및 조직 특이적 발현벡터를 개발하기 위하여, 우선 소의 위중 제4위 내의 미생물로부터 전체 유전자를 분리하였고, 이렇게 작성된 DNA library에서 섬유소분해 관련 유전자인 약 2.0 kb의 Cel D유전자를 크로닝하였으며, 췌장 특이적 발현 프로모터(rat elastase I: 약 200bp)를 크로닝한 후, 미세주입용 형질전환 재조합 벡터를 구축하기 위하여 rElastase I 프로모터 하류에 섬유소 분해 유전자(Cel D)를 연결하여 약 3.0 kb 크기의 재조합 벡터를 준비하였으며, 재조합 유전자를 1세포기 수정란 전핵내에 미세주입 하기 위해 Sal I과 BglII를 이용 유전자 단편을 만들었다. 구축된 유전자를 미세주입하기 위한 수정란을 회수하기위해 총68두의 돼지를 4-5두씩 분리사육하면서 발정동기화 및 과배란 유기를 위해 PG600, Altrenogest, FSH, hCG를 투여하였으며 hCG투여후 약54시간에 외과적 방법에 의해 총 1,359개의 수정란을 회수하였고, 이중 미세주입가능한 1세포기 수정란은 1,296개로 두당 평균 15.9개 였다. 1,296개의 1세포기 수정란 중에서 재조합 유전자(rE I-CelD)가 미세주입된 660개의 수정란을 32두의 수란돈에 외과적 방법에 의해 이식하였으며, 이식되어진 모돈 13두가 분만하여 40.6%의 임신율을 나타내었다. 이렇게 분만된 13두에서 총 65두(암:33두, 수:32두)의 자돈이 생산되었으며, 형질전환 여부를 판명하기 위해 자돈의 꼬리조직으로 부터 genomic DNA를 추출하고 PCR 검정을 실시하였다. PCR 검정 결과, 섬유소 분해 유전자가 도입된 자돈은 5두 이었으며, 그 결과를 Table에 나타내었다.(Table Omitted) Table 1 에서와 같이 섬유소 분해효소유전자가 형질전환된 자돈은 65마리 중 5마리로 7.69%의 형질전환율을 나타내었으며, 5마리의 자돈중 2두(암:1두, 수:1두)는 분만 후 즉시 폐사되었으며 2두(암:1두, 수:1두)는 86일령 그리고 14일령에 폐사하여 현재 1두(암)가 생존하여 섬유소 분해 사양 실험 중에 있다.

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Lumination of Epididymis and Electrophoretic Pattern of Proteins in Epididymal Fluid during Sexual Maruradon in Mouse (성적 성숙에 따른 생쥐 부정소의 강소형성과 부정소액내 단백질의 전기영동 양상)

  • 김문규;윤현수;최규완;윤용달
    • The Korean Journal of Zoology
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    • v.32 no.3
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    • pp.264-274
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    • 1989
  • In order to study the influence of spermatozoa and testicular fluid on the component and composition of proteins in epididymal fluid of mice, histological differentiation of testis and epididymis were observed during sexual maturation, and the proteins in epididymal fluids collected according to the characteristics of lumination were analyzed by electrophoresis (SDS-PAGE). In 10 day-old mouse, both of,testis and epididymis were undifferentiated. In 20 day-old mouse, epididymis was primitively luminated, but testis was not. In 35 day-old mouse, both of testis and epididymis were luminated and eaithdymal epithelium was differentiated into principal cells and clear cells. Spermatozoa were not transfered into epididymis yet. However, in 80 day-old mouse, both of festis and epididymis were fully differentiated and spermatozoa were transfered into epididymis. In electrophoretic paftem of proteins in epididymal fluid, a total of 28 kinds of proteins were identified, which were different from those of their sera. 12 kinds out of these proteins were epididymal specific protein(ESP) detected in epididymal fluid only, and the other 16 kinds(TEP) were also detected in testicular fluid. The proteins in epididymal fluid changed during sexual maturation and 3 kinds of the proteins changed quantitatively according to epididymal regions in adult. It may be concluded from the above results that the component and composition of the proteins in epididymal fluid changed by the influx of testicular fluid including spermatozoa into epididymis and regulation of the protein synthesis, secretion and/or absorption by the epididymal epithelium. Therefore it is strongly suggested that ESP and TEP in epididymal fluid play somehow significant roles on the maturation of epididymal spermatozoa.

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A Comparison Study of the Site Amplification Characteristics and Seismic Wave Energy Levels at the Sites near Four Electric Substations (4개 변전소시설 부지 인근관측소의 지반증폭 특성 및 파형에너지 수준 비교 연구)

  • Yoo, Seong-Hwa;Kim, Jun-Kyoung;Wee, Soung-Hoon
    • Journal of the Korean earth science society
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    • v.37 no.1
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    • pp.40-51
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    • 2016
  • The problem has been pointed out that the domestic design response spectrum does not reflect site amplification, particularly in the high frequency bands, including the fact that site specific response spectrum from the observed ground motions appears relatively higher than design response spectrum. Among various methods, this study applied H/V spectral ratio of ground motion for estimating site amplification. This method, originated from S waves and Rayleigh waves, recently has been extended to Coda waves and background noise for estimating site amplification. For limited time of periods, 4 electric substation sites had operated seismic stations at two separate locations (bedrock and borehole) within each substation site. H/V spectral ratio of S wave, Coda wave, and background noise, was applied to 36 accelerations of 3 macro earthquakes (Odaesan, Jeju and Gongju earthquakes), larger than magnitude 3.4. observed simultaneously at each bedrock location within 4 electric substation sites. Site amplifications at the bedrock location of 4 sites were compared among S wave, Coda wave energy, and background noise, and then compared to the previous results from the borehole location data. The site classification was also tried using resonancy frequency information at each site and location. The results suggested that all the electric substation sites showed similar site amplification patterns among S wave, Coda wave, and background noise. Each station showed its own characteristics of site amplification property in low, high and specific resonance frequency ranges. Comparison of this study to other results using different method can give us much more information about dynamic amplification of domestic sites characteristics and site classification.

Effect of the marginal position of prosthesis on stress distribution of teeth with abfraction lesion using finite element analysis (보철물 변연의 위치가 abfraction된 치아의 응력 분포에 미치는 영향에 대한 유한요소법적 분석)

  • Kim, Myeong-Hyeon;Lee, Cheong-Hee
    • The Journal of Korean Academy of Prosthodontics
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    • v.52 no.3
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    • pp.202-210
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    • 2014
  • Purpose: The aim of this study was to evaluate the stress concentration and distribution whether restoring the cavity or not while restoring with metal ceramic crown on tooth with abfraction lesion using finite element analysis. Materials and methods: Maxillary first premolar was selected and made a total of 10 finite element model. Model 1 was natural tooth; Model 2 was tooth with metal ceramic crown restoration which margin was positioned above 2 mm from CEJ; Model 3 was tooth with metal ceramic crown restoration which margin was positioned on CEJ; Model 4 was natural tooth which has abfraction lesion; Model 5 and 6 had abfraction lesion and the other condition was same as model 2 and 3, respectively; Model 7 was natural tooth which had abfraction lesion restored with composite resin; Model 8 and 9 was tooth with metal ceramic crown after restoring on abfraction lesion with composite resin; Model 10 was restored tooth on abfraction lesion with composite resin and metal ceramic crown restoration which margin is positioned on lower border of abfraction lesion. Load A and Load B was also designed. Von Mises value was evaluated on each point. Results: Under load A or load B, on tooth with abfraction lesion, stress was concentrated on the apex of lesion. Under load A or load B, on tooth that abfraction lesion was restored with composite resin, the stress value was reduced on the apex. Conclusion: In case of abfraction lesion was restored with composite resin, the stress was concentrated on the apical border of restored cavity regardless of marginal position. It was favorable to place crown margin on the enamel for restoring with metal ceramic crown.

A Study on the Simulation of Daily Precipitation Considering Spatial Probability Characteristics (공간적(空間的) 확률구조(確率構造)를 고려(考慮)한 일강수량(日降水量)의 모의발생(模擬發生)에 관한 연구(硏究))

  • Lee, Jae Joon;Lee, Won Hwan
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.6 no.3
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    • pp.31-42
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    • 1986
  • The probabilistic model was developed to give a spatial simlation of precipitation series to solve the problem of future need of water resources. The simulation of daily precipitation series at the sub-base stations was induced from the spatial structure of rainfall occurrence probability between the base station and the sub-base stations in the watershed. In this study Hadong was chosen as the base station in Seomjin river basin and Imsil, Boseong, Soonchang, Dongbok, and Gurye were also selected as the sub-base stations. The results of this study are as follows; 1) The separation technique of spatial precipitation state showed effectiveness in the spatial simulation method because the occurrence probability by each precipitation state (Wet-Wet, Dry-Wet, Wet-Dry, and Dry- Dry system) represented the stable value. 2) The daily precipitation series of the sub-base stations which were simulated from those of the base station showed that the simulated annual mean precipitations were similar to the observed data, but the precipitations in summer were decreased slightly. 3) The correlogram and power spectrum of the simulated monthly precipitation for the sub-base stations showed those of the observed sample with good agreement.

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Subpixel Shift Estimation in Noisy Image Using Iterative Phase Correlation of A Selected Local Region (잡음 영상에서 국부 영역의 반복적인 위상 상관도를 이용한 부화소 이동량 추정방법)

  • Ha, Ho-Gun;Jang, In-Su;Ko, Kyung-Woo;Ha, Yeong-Ho
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.47 no.1
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    • pp.103-119
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    • 2010
  • In this paper, we propose a subpixel shift estimation method using phase correlation with a local region for the registration of noisy images. Phase correlation is commonly used to estimate the subpixel shift between images, which is derived from analyzing shifted and downsampled images. However, when the images are affected by additive white Gaussian noise and aliasing artifacts, the estimation error is increased. Thus, instead of using the whole image, the proposed method uses a specific local region that is less affect by noises. In addition, to improve the estimation accuracy, iterative phase correlation is applied between selected local regions rather than using a fitting function. the restricted range is determined by analyzing the maximum peak and the two adjacent values of the inverse Fourier transform of the normalized cross power spectrum. In the experiments, the proposed method shows higher accuracy in registering noisy images than the other methods. Thus, the edge-sharpness and clearness in the super-resolved image is also improved.

The Effects of Regular Taekwondo Exercise on Brain wave activation and Neurotrophic Factors in Undergraduate male students (규칙적인 태권도 운동이 남자 대학생의 뇌파 활성화 및 뇌신경성장인자에 미치는 영향)

  • Kim, Young-Il;Ok, Duck-Pil;Cho, Su-Youn
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.2
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    • pp.412-422
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    • 2018
  • The purpose of this study was to investigate effects of 12-weeks aerobic exercise and taekwondo exercise on brain wave activation and brain-derived neurotrophic factors in undergraduate male students. Twenty four male subjects participated in this study. They were separated into a Control group(CG; n=8), Aerobic training group (ATG; n=8) and Taekwondo training group(TTG; n=8). ATG and TTG participated in Aerobic exercise training and Taekwondo exercise training for 12 weeks, 50~80 minutes per day, 3 times a week at 60~80% HRR respectively. All data were analyzed by repeated measures two-way ANOVA. As a result, there were no significant differences in the body composition and brain activation in all groups. However BDNF increased significantly after 12 weeks in the aerobic training groups(ATG). It is suggest that 12 weeks of regular Taekwondo exercise training did not statistically affect brain activation and neurotrophic factors in undergraduate students.

Relationships Between Bovine Growth Hormone Gene Polymorphism and Semen Characteristics in Hanwoo Bull (한우 종모우의 소 성장호르몬 유전자 다형과 정액성상과의 관계)

  • Lee, S.S.;Kim, J.H.;Jeong, J.;Park, N.H.
    • Journal of Animal Science and Technology
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    • v.44 no.6
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    • pp.693-700
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    • 2002
  • The objects of this study were to estimate gene frequencies of the bovine growth hormone(bGH), and to investigate the relationship between the bGH polymorphism and semen characteristics in Hanwoo bull. One hundred nine heads of Hanwoo bulls were used to identify bGH genotypes by the PCR-RFLP, followed by digestion with Alu I restriction enzyme. The frequencies of leucine(Leu) and Valine(Val) alleles were 0.88 and 0.12, respectively. Observed number of LL, LV and VV genotypes were 83, 25 and 1, respectively. Semen characteristics(semen volume, sperm concentration) were analyzed by GH genotypes in 25,559 ejaculates of 109 heads. Although bGH genotypes showed no significant effects on semen characteristics, those of bulls with VV genotype were tended to be lower than those of other bulls with LL or LV genotypes. And, in 1998, total sperm number(60.47${\times}$$10^8$) of VV bulls were significantly lower(P<0.05) than those(86.21${\sim}$92.22${\times}$$10^8$) of other genotypes bulls. This results provide that the VV bull in bGH locus may be worse, under the LL and LV bulls on semen characteristics. However, the number of examined VV bulls was only one and further investigations are needed to confirm the results.

Application of Silver Ion for Clinical Mastitis in Holstein Cows (임상형 유방염을 가진 홀스테인(Holstein) 소에 대한 은 이온의 적용 연구)

  • Seol, Jae-Won;Hur, Tai-Young;Jung, Young-Hun;Park, Sang-Youel;Kang, Seog-Jin
    • Journal of Veterinary Clinics
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    • v.27 no.3
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    • pp.246-251
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    • 2010
  • Mastitis is an inflammation of the mammary gland that develops as a response to pathogenic microorganisms. Somatic cell count (SCC) and bacteriological analysis are the most accepted tools to determine udder health. This study evaluated the effects of intra-mammary gland infusion of silver ion against clinical mastitis in Holstein cows. Silver ion (20 ${\mu}g$/ml) was infused through the intra-mammary gland in quarters having clinical mastitis and milk was collected to determine SCC, and levels of lactoferrin and bovine serum albumin (BSA). Silver ion infusion decreased udder inflammation, firmness and swelling, and reduced clots, BSA, lactoferrin and SCC in milk. However, milk yield and circulating blood cells remained unchanged. The silver ion-mediated reductions of BSA and SCC indicate reduced inflammation and bacterial activity in silver ion-treated mammary glands in Holsteins with mastitis, which may be exploited in a curative strategy.

Nuclear Remodeling and In Vitro Development of Bovine Oocytes Following Nuclear Transfer of Bovine Fetal Fibroblasts (태아 섬유아세포로 핵치환된 소 난자의 핵의 재구성과정과 체외 배발달)

  • Um, J. H.;S. J. Uhm;Kim, N-H;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.1
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    • pp.59-67
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    • 2000
  • This study was investigated the developmental potential of bovine embryos following nuclear transfer with bovine fetal fibroblasts (BFF). BFF were isolated from a male 45-day-old-fetus. Non-starved BFF labeled with MitoTracker were transferred into perivitelline space of enucleated oocytes. BFF-oocyte units were fused by electric pulse, and then fused oocytes were activated with calcium ionophore A23187 and subsequently 6-dimethylaminopurine (6-DMAP). The resulting zygotes were placed into CRlaa bovine embryo culture medium. Transfer of the nucleus into enucleated oocyte led to premature chromosome condensation, swelling and pronucleus formation. Remodeled oocytes were developed to the mitotic and 2-cell stage at 18 to 26 h after nuclear transfer. The incidence of in vitro development to the blastocyst stages was 21% of fused oocytes. Mitochondria of BFF eliminated rapidly and were not detected at 8 h after fusion. These results suggest that BFF can be successfully reprogrammed in enucleated bovine oocytes, and that reconstructed embryos can develop to the blastocyst stage.

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