• Title/Summary/Keyword: CgGH

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CgGH insertion functional domain analysis in transgenic G1 and G2 and G3 mutiara catfish (Clarias gariepinus) broodstock

  • Buwono, Ibnu Dwi;Grandiosa, Roffi;Mulyani, Yuniar
    • Fisheries and Aquatic Sciences
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    • v.25 no.1
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    • pp.1-11
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    • 2022
  • Catfish is one of the most important freshwater fish farming commodities in Indonesia. Higher catfish production can be achieved by cultivating transgenic catfish carrying the growth hormone (GH) gene of African catfish (Clarias gariepinus GH, CgGH). This research focuses on analysis of the presence of the CgGH gene in transgenic G1, G2, and G3 mutiara catfish broodstock, as an indication of stable CgGH inheritance. CgGH gene was isolated using the RNeasy mini kit and RT-PCR. RT-PCR revealed amplicons measuring approximately 600 bp in transgenic G0, G1, G2, and G3 mutiara catfish. The CgGH consensus sequence similarities ranged from 93.76% to 97.06%, with four functional domain sites (somatotropin-1, somatotropin-2, four α-helix, N-glycosylation, four cysteine residues) of fish GH proteins. The functional domains of fish GH proteins are conserved in G1, G2, and G3 and indicate stable exogenous GH inheritance to produce transgenic catfish strains in each generation.

Effect of Pronuclear Injection with Human Growth Hormone Gene on Development and PCR-Screening in Rabbit Embryos (사람성장호르몬 유전자의 전핵내 미세주입이 토끼 수정란의 체외발달에 미치는 영향과 PCR검색)

  • Kang, T. Y.;Chae, Y. J.;Lee, H.;Lee, K. K.;Park, C. S.;Lee, H. J.
    • Journal of Embryo Transfer
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    • v.13 no.2
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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.

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Evaluation of Sleep Comfort for Indoor Thermal Environment based on the Physiological Signal Analysis (생리신호 분석을 이용한 실내온열환경에서의 수면 쾌적성 평가)

  • 이낙범;임재중;금종수;이구형;최호선
    • Science of Emotion and Sensibility
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    • v.3 no.2
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    • pp.75-84
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    • 2000
  • 문명의 발달과 함께 수면부족으로 인한 여러 가지 스트레스와 질환이 증가하게 되어 최근 수면연구에 대한 관심이 증가하고 있다. 본 연구는 다양한 온열환경 조건에서의 쾌적한 수면을 위한 온열환경을 제시하기 위해, 5명의 여성 피험자를 대상으로 22$^{\circ}C$, 26$^{\circ}C$, 3$0^{\circ}C$의 일정온도 조건과 $25^{\circ}C$에서 1시간 후와 2시간 후에 각각 1, 2$^{\circ}C$를 상승시켜주는 변동온도 조건하에서 수면생리신호를 측정하였다. 그리고, 수면단계 평가를 이용하여 총 수면시간, 깊은 수면의 비율, 그리고 최초 수면시작 시간에서 최초의 서파 수면이 나타나기까지의 지연시간 등의 수면효율을 평가하였다. 그 결과, 일정온도 조건에서는 26$^{\circ}C$에서 총 수면시간(466.7$\pm$10.25분)과 깊은 수면의 비율(33.1$\pm$4.95%)은 타 조건에 비해 높게 나타났고, 최초 서파수면까지의 지연시간(9.8$\pm$3.33분)은 타 조건에 비해 낮게 나타나 쾌적한 수면을 위한 가장 적절한 온열조건임을 관찰할 수 있었다. 그리고 변동온도 조건에서는 4가지 온열조건간에는 큰 차이가 나타나지 않았지만, 모든 조건에서 일정온도 조건보다는 좋은 결과를 나타내었다. 또한 수면 중 신체 움직임과 설문 분석에서도 동일한 결과를 보였다. 본 연구를 통해, 수면생리신호를 이용한 수면 쾌적성 분석은 수면의 질적인 상태를 관찰하는데 매우 적합한 파라메터를 도출할 수 있으며, 여러 가지 수면환경 조건을 평가하는데 매우 유용한 지표가 될 수 있음을 보였다.e results suggest that hCG treatment at 7 days after insemination could be used to increase the pregnancy rate of embryo transfer, and transfer, and only the recipients with PUN concentration of <12 mg/dl were influenced by treatment with hCG./TEX>이었으며, 이는 화성 기원을 지시한다.sucrose를 이용한 2단계 희석이 수정란의 생존성을 향상시키는 것으로 나타났다. 또한 발육 단계별 생존성에 있어서는 발육이 진전된 확장배 반포 시에 동결하는 것이 배반포기에 동결하는 것 보다 유리한 것으로 나타났다.ody를 사용하여 flow cytometery해석을 실시하는 한편 125I-hGH binding assay에 의하여 hGH binding activity를 측정하였다. 최종적으로 GH signal transduction의 target genedf으로 알려져 있는 serine protease inhibitor 2.1(Spi 2.1) gene의 promotor activity를 검토한 결과 hGHR을 transfect한 CHO Cell에 있어서 hGH의 농도에 의존적으로 증가되었다. 따라서 본 실험에서 cloning한 cDNA hGHR는 native hGHR와 같은 기능을 가지는 것으로 판명되었다.것으로 판명되었다..ments of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to

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Increased mRNA Related Ovarian Maturation during Induction of Maturational Competence in Red Seabream, Pagrus major (참돔, Pagrus major의 성숙능력 유도시 증가된 난성숙 관련 mRNA)

  • Choi, Cheol-Young;Chang, Young-Jin;Takashima, Fumio
    • Development and Reproduction
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    • v.4 no.1
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    • pp.125-131
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    • 2000
  • This study has used differential display-PCR, to amplify genes transcribed during the ovarian maturation induced by human chorionic gonadotropin (hCG). The cDNA expressed at the times of acquisition of oocyte maturational competence in red seabream (Pagrus major) following treatment with hCG was amplified and cloned. A full-length of cDNA for p. major was isolated using differential display-PCR and 5'RACE. This cDNA clone contained 2,662 nucleotides including the open reading frame that encoded 434 amino acids. Homology analyses, using the GenBank and EMBL general database searches, indicated that the nucleotides sequence of the cDNA does not have high homology with any other genes. This cDNA was judged to be a gene, which induction of maturational competence coincides with increase of mRNA related ovarian maturation. Consensus sequences which were consistent with protein kinase C phosphorylation sites and casein kinase II phosphorylation sites were identified. in vitro, the transcription level of mRNA related ovarian maturation increased between 9hr and 24hr following treatment of ovarian follicles with hCG. It was also increased after GtH-II (300 ng/ml) stimulation. Furthermore, in vivo, mRNA related ovarian maturation was rarely expressed prior to the acquisition of oocyte maturational competence, but was strongly expressed after the acquisition of oocyte maturational competence, suggesting that the hCG induction of maturational competence is brought about by the de novo synthesis of the mRNA related ovarian maturation in p. major.

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Effects of intrauterine growth restriction during late pregnancy on the cell growth, proliferation, and differentiation in ovine fetal thymuses

  • Zi, Yang;Ma, Chi;He, Shan;Yang, Huan;Zhang, Min;Gao, Feng;Liu, Yingchun
    • Animal Bioscience
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    • v.35 no.7
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    • pp.989-998
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    • 2022
  • Objective: This study investigated the effects of intrauterine growth restriction (IUGR) during late pregnancy on the cell growth, proliferation, and differentiation in ovine fetal thymuses. Methods: Eighteen time-mated Mongolian ewes with singleton fetuses were allocated to three groups at d 90 of pregnancy: restricted group 1 (RG1, 0.18 MJ ME/body weight [BW]0.75/d, n = 6), restricted group 2 (RG2, 0.33 MJ ME/BW0.75/d, n = 6) and control group (CG, ad libitum, 0.67 MJ ME/BW0.75/d, n = 6). Fetuses were recovered at slaughter on d 140. Results: The G0/G1 phase cell number in fetal thymus of the RG1 group was increased but the proliferation index and the expression of proliferating cell nuclear antigen (PCNA) were reduced compared with the CG group (p<0.05). Fetuses in the RG1 group exhibited decreased growth hormone receptor (GHR), insulin-like growth factor 2 receptor (IGF-2R), and their mRNA expressions (p<0.05). For the RG2 fetuses, there were no differences in the proliferation index and PCNA expression (p>0.05), but growth hormone (GH) and the mRNA expression of GHR were lower than those of the CG group (p<0.05). The thymic mRNA expressions of cyclin-dependent protein kinases (CDKs including CDK1, CDK2, and CDK4), CCNE, E2-factors (E2F1, E2F2, and E2F5) were reduced in the RG1 and RG2 groups (p<0.05), and decreased mRNA expressions of E2F4, CCNA, CCNB, and CCND were occurred in the RG1 fetuses (p<0.05). The decreased E-cadherin (E-cad) as a marker for epithelial-mesenchymal transition (EMT) was found in the RG1 and RG2 groups (p<0.05), but the OB-cadherin which is a marker for activated fibroblasts was increased in fetal thymus of the RG1 group (p<0.05). Conclusion: These results indicate that weakened GH/IGF signaling system repressed the cell cycle progression in G0/G1 phase in IUGR fetal thymus, but the switch from reduced E-cad to increased OB-cadherin suggests that transdifferentiation process of EMT associated with fibrogenesis was strengthened. The impaired cell growth, retarded proliferation and modified differentiation were responsible for impaired maturation of IUGR fetal thymus.

Cloning of MT -hGH Gene-injected Rabbit Embryos by Nuclear Transplantation (사람성장호르몬 유전자주입 토끼수정란의 핵이식에 의한 복제)

  • Kang, T.Y.;Chae, Y.J.;Lee, H.;Park, C.S.;Lee, H.J.
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.419-424
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    • 1998
  • The present study was carried out to examine the efficiency of cloning of transgenic embryos by nuclear transfer(NT) using gene-injected rabbit embryos. The rabbit embryos at pronuclear stage were microinjected with methallothionein-human growth hormone(MT-hGH) gene and cultured to 8- and 16-cell in TCM-199 containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% $CO_2$incubator. The recipient oocytes were collected from the oviducts 14~16 h after hCG injection. The oocytes were enucleated and activated with 5$\mu$M ionomycin and 2mM 6-dimethylaminopurine. Blastomeres form gene-injected embryos were transferred into the enucleated oocytes by micromanipulation. The nuclear transplant oocytes were electrofused and co-cultured with rabbit oviductal cells. Following 120 h of culture, blastocysts were prepared for gene analysis by polymerase chain reaction(PCR). In previous experiment, the rate of gene-positive embryos detected by the nested PCR analysis was significantly decreased while developing to blastocyst(25%)(Kang et al., 1998). The fusion rate of gene-injected blastomeres was significantly(P<0.05) lower than non-injected blastomeres(66% vs 80%). However, the NT embryos that were derived from gene-injected donor embryos did not differ from control embryos in development to the blastocyst stage(39% vs 31%). Of the 43 NT blastocysts developed from the gene-injected donor embryos, twelve(28%) were positive for the injected DNA. The results indicate that NT with gene-injected embryos can be successfully used for cloning and multiplication of transgenic embryos, furthermore applicable to improvement of transgenic animal production.

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Effect of Aerobic Exercise on Factors Relative to the Brain Nerve Growth in Girls (유산소운동이 여자 아동의 뇌세포 생성 관련인자에 미치는 영향)

  • Pyun, Mi-Young;Cho, Han-Sam;Jeon, Jae-Young;Kim, Jong-Won;Lee, Kyung-Hee;Lim, Chun-Kyu;Kim, Tae-Un;Kim, Hyun-Jun;Kwak, Yi-Sub;Ko, Ki-Jun
    • Journal of Life Science
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    • v.20 no.6
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    • pp.948-954
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    • 2010
  • Exercise can favorably influence brain plasticity by facilitating neurogeneration, neuroadaptivity, and neuroprotection. Aerobic exercise has been reported to change brain nerve growth factors (growth hormone, insulin like factor-1, estrogen and serotonin). The purpose of this study was to demonstrate the effects of aerobic exercise for 12 weeks on brain nerve growth factors in girls. Fourteen female participants in elementary school grades 1 through 3 were randomly allocated to the exercise group (EG, n=6) and control group (CG, n=8). The EG participated in 60 minutes of modified ballet exercise as aerobic training three days a week for 12 weeks. Based on comparison between groups by two-way ANOVA with repeated measures, aerobic exercise program participants experienced decreased weight (p<0.01), BMI (p<0.01), fat mass (p<0.001), fat percent (p<0.001) and increased LBM (lean body mass) percent (p<0.001). In addition, we detected that aerobic exercise decreased the level of serotonin (p<0.05) and increased the level of GH (p<0.05) and IGF-1 (p<0.05). These findings suggest that aerobic exercise programs can be an efficient intervention to change body composition, alleviate central fatigue, improve brain function, and induce brain cell proliferation in girls.