• Title/Summary/Keyword: Clone animal

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Variation of Mitochondrial DNA Restriction Fragments of Common Rats, Rattus norvegicus caraco Pallas (Mammalia , Redentia) , from Cheongju , Korea (청주에 서식하는집쥐[Rattus norvegicus caraco Pallas(설치목, 포유강)]의 미토콘드리아 DNA 절단단편의 변이)

  • Hung Sun Koh;Yong Seok Roh;Sang Bok Kim;Byung Sun Yoo
    • Animal Systematics, Evolution and Diversity
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    • v.11 no.4
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    • pp.409-416
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    • 1995
  • Forty samples of common rats (Rattus norvegicus caraco) from Cheongu, Korea, were used for the analyses of mitochondrial DNA (mtDNA) fragment patterns resulted from the digestion with eight restriction enzymes. A total of 36 fragments were recognized and six mtDNA clones were revealed . The nucleotide-sequence divergences (p) among six mtDNA clones ranged from 0.35% to 2.73%. moreover, the six clones were grouped into three major subgroups ; the first, second , and third subgroup were composed of 29 samples of three clones, ten samples of two clones, and one sample of one clone, respectively. The second and third subgroups were different in their mtDNa genotype of Pvu II from the first subgroup, and the third subgroup differed in the genotype of Dra I from other two subgroups. Futhermore, the maximum divergence among common rats from Korea in this study is greater than that among common rats from the United States and Japan by Brown and Simpson (1981). Further analyses with additional sample from other localities in Korea appeared to be necessary in order to clarify the taxnomic status of the distinct mtDNA subgroups.

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Isolation and Characterization of the Ribosomal Protein 46 Gene in Drosophila melanogaster

    • Animal cells and systems
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    • v.2 no.1
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    • pp.113-116
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    • 1998
  • A cDNA clone coding for ribosomal protein 46 (rp46) which is a component of 60S ribosomal large subunit has been identified from Drosophila melanogaster. A cDNA clone encoding S. cerevisiae rp46 was used as a probe to screen a Drosophila larvae cDNA library. The DNA sequence analysis revealed that the cDNA coding for Drosophils rp46 contains a complete reading frame of 153 nucleotides coding for 51 amino acids. The deduced amino acid sequence showed 71-75% homology with those of other eukaryotic organisms. Northern blot analysis showed that about 1-kb rp46 transcripts are abundant throughout fly development. Whole mount embryonic mRNA in situ hybridization also showed no preferential distribution of the transcripts to any specific region. The chromosomal in situ hybridization revealed that the identified gene is localized at position 60C on the right arm of the second polytene chromosome with a possibility of single copy.

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Isolation and Characterization of Phosphorus Accumulating Microorganisms under Liquid Fertilization of Swine Slurry (액비화 과정 중 인 이용 우수미생물 분리 및 특성)

  • Lim, Joung-Soo;Cho, Sung-Back;Hwang, Ok-Hwa;Yang, Seung-Hak
    • Journal of Animal Environmental Science
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    • v.20 no.2
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    • pp.77-84
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    • 2014
  • This study was conducted to investigate the bacterial capability to accumulate phosphorus during liquid composting process of pig slurry. Storage liquid compost and pig slurry were analyzed by using MALDI-TOF technique, which showed the colonies of Acinetobacter towneri and Bacillus licheniformis. In addition, bacterial colonies were isolated under high phosphoric acid conditions using X-phosphate MOPS medium with the addition of 2 mM $K_2HPO_4$. Microbial growth was observed in high and low phosphoric conditions due to the growth of bacterial diversities in the liquid fertilizer and slurry. The colonies isolated in the high phosphoric acid medium were uncultured bacterium clone and Acinetobacter sp. were identified by analysis of 16S rRNA gene sequences. Uncultured bacterium showed higher growth rate and excellent phosphorus ability then Acinetobacter sp.. In addition to Paenibacillus sp. AEY-1 isolated from pig slurry performed excellent phosphorus utilizing capability.

Embryonic Stem Cell and Nuclear Transfer

  • 임정묵
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.06a
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    • pp.19-25
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    • 2002
  • Researches on manipulation pluripotent stem cells derived from blastocysts or promordial germ cells (PGCs) have a great advantages for developing innovative technologies in various fields of life science including medicine, pharmaceutics, and biotechnology. Since the first isolation in the mouse embryos, stem cells or stem cell-like colonies have been continuously established in the mouse of different strains, cattle, pig, rabbit, and human. In the animal species, stem cell biology is important for developing transgenic technology including disease model animal and bioreactor production. ES cell can be isolated from the inner cell mass of blastocysts by either mechanical operation or immunosurgery. So, mass production of blastocyst is a prerequisite factor for successful undertaking ES cell manipulation. In the case of animal ES cell research, various protocol of gamete biotechnology can be applied for improving the efficiency of stem cell research. Somatic cell nuclear transfer technique can be applied to researches on animal ES cells, since it is powerful tool for producing clone embryos containing genes of interest. In this presentation, a brief review was made for explaining how somatic cell nuclear transfer technology could contribute to improving stem cell manipulation technology.

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SEQUENCE ANALYSIS AND COMPARISON OF BOVINE αS1-CASEIN GENOMIC DNA

  • Lin, C.S.;Huang, M.C.;Choo, K.B.;Tseng, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.4
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    • pp.541-547
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    • 1993
  • A phage clone containing the partial ${\alpha}_{S1}$-casein gene was isolated from a bovine genomic library by using mixed probes of ovine ${\alpha}_{S1}$-, ${\beta}$- and ${\kappa}$-casein cDNAs. Restriction enzyme mapping analysis for 14.6 kb revealed that the map was in conflict with the report of Meade et al. (1990), especially in the 3'-end fragment. Sequence analysis of 12.6 kb revealed a high AT/GC ratio (1.64); we have identified eight exon sequences according to the bovine ${\alpha}_{S1}$-casein cDNA sequence. The same exon/intron splice junction sequence was observed between these exons. We suggest that the bovine ${\alpha}_{S1}$-casein gene night contain a minimum of 18 exons and the full length is approximately 18-19 kb.

Molecular Cloning, Characterization, and Expression Analysis of Chicken Δ-6 Desaturase

  • Kang, Xiangtao;Bai, Yichun;Sun, Guirong;Huang, Yanqun;Chen, Qixin;Han, Ruili;Li, Guoxi;Li, Fadi
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.1
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    • pp.116-121
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    • 2010
  • Long-chain polyunsaturated fatty acids (LC-PUFA) promote the development of brain and vision of the fetus, relieve inflammation, inhibit oral dysplasia of rumor cell, decrease the incidence of cardiovascular disease and regulate arrhythmia. ${\Delta}-6$ desaturase is the rate-limited enzyme in the desaturation process. This study reports the cloning, characterization and tissue expression of a ${\Delta}-6$ desaturase gene in the chicken. PCR primers were designed based on the predicted sequence of chicken ${\Delta}-6$ desaturase (accession number: XM421053) and used to isolate a cDNA fragment of 1,323 bp from chicken liver. Based on the 1,323 bp fragment an EST (BI390105) was obtained by BLAST. The EST and 5'nd of the 1,323 bp fragment were partially overlapped. Gene specific primers derived from the EST were used for amplification of the 5'nd. Another gene-specific primer derived from the 1,323 bp fragment was used for amplification of the 3'nd by 3'ACE. Then the three overlapping cDNA sequences obtained were assembled with DNAMAN software and a full-length ${\Delta}-6$ desaturase of 2,153 bp was obtained. The full-length cDNA contained an ORF of 1,335 bp with a 5'ntranslated region of 147 nucleotides followed by an ATG initiation codon. Stop codon TGA was at position 1,481-1,483 bp. The deduced amino acids shared an homology above 77% with bovine, mice, orangutan, rat and human. The protein sequence had three histidine-rich regions HDFGH (HisI region), HFQHH (HisII region) and HH (HisIII region), a cytochrome $b_{5}$-like domain containing a heme-binding motif and two transmembrane domains. Sequence analysis of the chicken genomic DNA revealed that the coding sequence of chicken ${\Delta}-6$ desaturase included 12 exons and 11 introns. Semi-quantitative RT-PCR showed that the ${\Delta}-6$ desaturase expression levels were in turn liver, spleen, pancreas, lung, breast muscle, heart, and abdominal fat. The expression of ${\Delta}-6$ desaturase in liver was significantly higher than that in breast muscle (p<0.01). The expression of ${\Delta}-6$ desaturase in lung was significantly higher than that in abdominal fat (p<0.01). This is the first clone of chicken ${\Delta}-6$ desaturase.

Phylogenetic Analysis of 16S rDNA Sequences Manifest Rumen Bacterial Diversity in Gayals (Bos frontalis) Fed Fresh Bamboo Leaves and Twigs (Sinarumdinaria)

  • Deng, Weidong;Wanapat, Metha;Ma, Songcheng;Chen, Jing;Xi, Dongmei;He, Tianbao;Yang, Zhifang;Mao, Huaming
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1057-1066
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    • 2007
  • Six male Gayal (Bos frontalis), approximately two years of age and with a mean live weight of $203{\pm}17$ kg ($mean{\pm}standard\;deviation$), were housed indoors in metabolism cages and fed bamboo (Sinarundinaria) leaves and twigs. After an adjustment period of 24 days of feeding the diet, samples of rumen liquor were obtained for analyses of bacteria in the liquor. The diversity of rumen bacteria was investigated by constructing a 16S rDNA clone library. A total of 147 clones, comprising nearly full length sequences (with a mean length of 1.5 kb) were sequenced and submitted to an on-line similarity search and phylogenetic analysis. Using the criterion of 97% or greater similarity with the sequences of known bacteria, 17 clones were identified as Ruminococcus albus, Butyrivibrio fibrosolvens, Quinella ovalis, Clostridium symbiosium, Succiniclasticum ruminis, Selenomonas ruminantium and Allisonella histaminiformans, respectively. A further 22 clones shared similarity ranging from 90-97% with known bacteria but the similarity in sequences for the remaining 109 clones was less than 90% of those of known bacteria. Using a phylogenetic analysis it was found that the majority of the clones identified (57.1%) were located in the low G+C subdivision, with most of the remainder (42.2% of clones) located in the Cytophage-Flexibacter-Bacteroides (CFB) phylum and one clone (0.7%) was identified as a Spirochaete. It was apparent that Gayal have a large and diverse range of bacteria in the rumen liquor which differ from those of cattle and other ruminants. This may explain the greater live weights of Gayal, compared to cattle, grazing in the harsh natural environments in which Gayal are located naturally.

The Status of Laboratory Animal Production and Visions in the 21st Century - Review -

  • Gartner, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.7
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    • pp.1142-1151
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    • 1999
  • Today, laboratory animal production has decreased world-wide to half the number estimated in 1970 of more than 100 Mio. This is due to the cell-biological assays which replaced animal experimentation as a first allround method to solve biomedical problems. Animal experimentation remains the most significant experimental method for the study of higher organized physiological systems and their multifactorial connections. This requires maximal uniformity of all quantitative traits among the animals used for such studies (mainly mice and rats) and stability of these traits for reproducing such studies at any time world-wide. The success of the developed methods for the standardization of laboratory animals was analyzed and were found only partly be acceptable. Getting a higher degree of uniformity among standardized inbred animals is blocked by "intangible variance". This is caused by influences of ooplasm, shown by experimental twin and clone studies. Manipulation of this component of variance is essential in the future. - Genetic drifts impair the necessary stability of biological traits. There are a few disadvantages associated with the cryopreservation of embryos and other methods are required. - Dogs and cats were replaced by pigs as laboratory animals. A new line of animal production will evolve over the next 25 years with similarities to the present laboratory animal production, because in future pigs were used as donors for xenotransplants for men.

Cloning and Characterization of Bovine 5-Cytosine DNA Methyltransferase I cDNA

  • Lee, Poongyeon;Min, Kwan-Sik;Lee, Hyun-Gi;Kim, Soon-Jeung;Chung, Hee-Kyoung;Seo, Myung-Kyu;Lee, Yun-Keun;Kim, Sung-Woo;Park, Jin-Ki
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.39-39
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    • 2003
  • Methylation of DNA 5-cytosine in mammalian early embryo affects great deal in nuclear reprogramming and chromatin remodeling of developing embryo. Current efforts to clone and produce cloned animals including transgenic animals face various problems including low birth rate, irregular development, and so on. In this report, cDNA for the one of house keeping methyltransfcrase, Dnmt1 was cloned from bovine somatic tissues and was analyzed for its nucleotide sequences to investigate the structure and function of the gene in bovine early development. Nucleotide sequence of bovine Dnmt1 homologue showed 76.8% identity with that of human Dnmtl and 66.4% with mouse Dnmt1. Translated amino acid sequence showed 88.4% homology with human homologue and 75.8% homology with mouse counterpart. Three types of Dnmt1 are reported in mouse and human, and are likely present in bovine tissues. Understanding of role of Dnmt1 in bovine development may shed a light in the field of animal, especially bovine cloning.

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Production of Clone Animals by Nuclear Transplantation I. Effects of Electrostimulation on Membrane Fusion of Embryos and Activation of Oocytes in Mouse (핵치환에 의한 Clone Animal의 생산에 관한 연구 I. 생쥐 수정란의 세포막 융합과 난모세포의 활성화에 미치는 전기자극의 효과)

  • 이상진;구덕본;이상민;박흠대;정순영;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.3
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    • pp.217-228
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    • 1994
  • These experiments were carried out to establish the optimal condition of electrostimulatin inducing cell fusion and oocyte activation for nuclear transplantation in mouse embryos. Eggs selected for cell fusion or activation by electrostimulation were equilibrated for 5~10 min. in 0.3M sucrose solution and electrostimulated for 60$\mu$sec using 1 pulse of 60, 70, 80, 90 or 100 volts DC with electrodes 0.2 mm apart. Then they were cultured in 20${mu}ell$ dropsof Tyrode's solution. The results of these experiments are as follows : 1. When one pulse of 60, 70, 80, 90 or 100 volts DC for 60$\mu$sec were applied to 2-cell embryos for fusion of blastomeres, fusion rates were 50.0, 81.7, 91.7, 100 and 100%, respectively ; and developmental rates of fused embryos to blastocyst were 76.7 to 81.5%. Higher fusion rates were observed in 90V and 100V. 2. The average cell number in fused embryos developed to blastocyst was about half of the cell number in diploid controls; and the cell number decreased with increasing of voltages. 3. When pulse numbers were increased, fusion rates improved, but developmental rates were not signficiantly different from the group for which the number of pulse was not increased. And the cell number of blastocyst decreased even more. 4. Oocytes aged for 6hrs after ovulation were electrostimulated for oocyte activation by the same method used for cell fusion. Rates of oocyte activated by electrostimulation were 45.3 to 60.4%, and fragmentation rates were 7.5~15.1%. The lysis rates were 17.0~34.0%. The results of these experiments indicate that the optimal condition for achieving cell fusion and activation is 1 pulse, duration 60$\mu$sec in 90 Volt. The results also show that this condition is suitable for nuclear transplantation using mouse eggs.

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