• Title/Summary/Keyword: DnaJ

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Expression of GFP Gene in Porcine Embryos after ICSI with Different DNA Binding Methods

  • Han, J. H.;Kim, S.W.;Lee, Y.K.;Lee, P.Y.;Park, C.G.;Lee, S.E.;Baek, K.N.;Lee, H.G.;Lee, J.Y.;Chang, W.K.;Park, J.K.
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.191-191
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    • 2004
  • Transgenic animals are produced primarily by microinjecting exogenous DNA into the male pronuclei of a zygote. Microinjection method for gene transmitting is successful in mice but not efficient in farm animals, limiting it's general utility such as a large scale facility and labour. Based on our finding that sperm cells bind with exogenous DNA, sperm was used as a vector for producing transgenic animals to introduced green fluorescence protein(GFP) gene. (omitted)

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Investigation of Deletion Variation and Methylation Patterns in the 5' LTR of Porcine Endogenous Retroviruses

  • Jung, K.C.;Simond, D.M.;Moran, C.;Hawthorne, W.J.;Jeon, J.T.;Jin, D.I.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.11
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    • pp.1572-1575
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    • 2008
  • The xenotransplantation of pig organs and cells can be related with a risk of transmission of infectious diseases to human. Previous findings indicate that the regulatory region of PERV for retroviral transcription, replication and integration into the cellular DNA is located on the 5' Long Terminal Repeat (LTR). The objective of this study is the investigation of methylation and deletion status of the PERV 5' LTR region which can be used for regulating PERV expression. We compared the sequences of genomic DNA and bisulfite-treated genomic DNA from PK-15 cells expressing PERV to observe the methylation status of the 5' LTR. Our results showed that the CpG sites of U3 were methylated and methylation was inconsistent in the R and U5 regions. Also, variable numbers of 18 bp repeats and 21 bp repeats were detected on 5' LTR by sequencing analysis. The consistent U3 methylation might be indicative of host suppression of expression of the retroviruses.

Extension of a 5'- or 3'-end Genomic DNA Sequence by a Single PCR Amplification

  • Jeon, Taeck J.
    • Journal of Integrative Natural Science
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    • v.1 no.3
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    • pp.230-233
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    • 2008
  • A simple and rapid method is described for extending the 5'- or 3'-end genomic sequence of a known partial sequence by only a single round of PCR. This method involves digesting and ligating genomic and plasmid DNAs, and amplifying the 5'-upstream or 3'-end downstream sequence of the known DNA sequence, using two primers, one gene specific and the other plasmid specific. A single round of PCR amplification is sufficient to produce gene-specific bands detectable in gels. By using this approach, 5'-end genomic sequence of the D-amoeba sams gene was extended.

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Ancestral Remnants in the Deoxyribonucleic Acid from Pseudomonas (Deoxyribonucleic Acid 속의 Pseudomonas로부터의 Ancestral Remnants)

  • J. De Ley;In Won Park
    • Journal of the Korean Chemical Society
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    • v.13 no.4
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    • pp.355-364
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    • 1969
  • Cross-hybridizations between DNA of two pseudomonads and a xanthomonad suggested that the three DNA types had a considerable section in common. The existence of this common part was proved by hybridization of preselected DNA, i.e. DNA resulting from a previous hybridization between any one set of two DNA types, with the third type. It was thus shown that about 50% of the DNA of the three organisms was similar. This common part was isolated in pure state and its % (G+C) was found to be indentical to the overall base composition of the native DNA. The evolutionary drift in % (G+C) could thus not be detected. The total molecular weight of the chromosornal DNA/bacterial nucleoid was determined to be 2.4 ${\times} 10^9$daltons. It can therefore be estimated that the common putida-fluorescenspelargonii DNA part consists of some 2,000 cistrons. P. putida and P. fluorescens share an additional 1,300 cistrons, and all xanthomonads share at least an additional 1,000 cistrons.

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Genetic Variation of Mitochondrial DNA in Duroc (Sus Scrofa) Using Single Stranded Conformation Polymorphism Analysis (Single Stranded Conformation Polymorphism 분석에 의한 돼지 Duroc 품종의 미토콘드리아 DNA 유전적 변이)

  • Cho, I.C.;Jung, Y.H.;Jung, J.K.;Seong, P.N.;Kim, B.W.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.911-916
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    • 2003
  • The mitochondrial DNA(mtDNA) D-loop region was amplified from Duroc(Sus scrofa) by polymerase chain reaction(PCR). The oligonucleotide primer used to amplify the Sus scrofa mtDNA D-loop region was designed using tRNA-Pro and tRNA-Phe sequence in mtDNA regions highly conserved in many other animal species. There were 1,145 base pairs(bp) in the D-loop region. The middle of the region contained 10 tandem repeat of an 10-bp Sus scrofa-specific sequence, TACACGTGCG. We designed primers for PCR-mediated single stranded conformation polymorphism(SSCP) analysis that amplified a 345 bp fragment, which contained the most variable region according to our sequencing data. SSCP analysis of denatured amplification products was carried out by polyacrylamide(8%) gel electrophoresis followed by ethidium bromide staining. The SSCP analysis identified two band patterns(A and B) and comparision of these two nucleotide sequences identified 21 base substitutions. These results show that SSCP analysis of the D-loop region is useful for detecting the genetic polymorphism.

Application of Breed-specific DNA Markers for the use of Identifying Major Pure Pig Breeds Maintained in Korea (대한민국내 주요 돼지 품종의 순종 식별을 위한 품종특이 DNA marker의 활용)

  • Seo, B.Y.;Kim, J.H.;Park, E.W.;Lim, H.T.;Cho, I.C.;Kim, B.W.;Oh, S.J.;Cheong, I.C.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.735-742
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    • 2004
  • This study was conducted for the identification of pure Landrace, Large White and Duroc breeds which are mainly maintained in Korea using DNA markers. We used known KIT and MC1R mutations, which were related coat color in pigs, and pig mitochondrial DNA variations. The KIT mutation was used to distinguish white and colored animals. Duroc breed could be discriminated from other colored breeds using the MC1R mutation N121D. Discriminating Landrace and Large White was possible using the l l-bp duplication of D-Ioop region and alternative initiation codon of ND2. In conclusion, identification of Landrace, Large White and Duroc breeds was might be possible using the procedure designed in this study.

Molecular Characterization and Expression Pattern of Gene IGFBP-5 in the Cashmere Goat (Capra hircus)

  • Wang, X.J.;Shi, J.J.;Yang, J.F.;Liang, Y.;Wang, Y.F.;Wu, M.L.;Li, S.Y.;Guo, X.D.;Wang, Z.G.;Liu, D.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.606-612
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    • 2012
  • Insulin-like growth factor-binding protein-5 (IGFBP-5) is one of the six members of IGFBP family, important for cell growth, apoptosis and other IGF-stimulated signaling pathways. In order to explore the significance of IGFBP-5 in cells of the Inner Mongolian Cashmere goat (Capra hircus), IGFBP-5 gene complementary DNA (cDNA) was amplified by reverse transcription polymerase chain reaction (RT-PCR) from the animal's fetal fibroblasts and tissue-specific expression analysis was performed by semi-quantitative RT-PCR. The gene is 816 base pairs (bp) in length and includes the complete open reading frame, encoding 271 amino acids (GenBank accession number JF720883). The full cDNA nucleotide sequence has a 99% identity with sheep, 98% with cattle and 95% with human. The amino acids sequence shares identity with 99%, 99% and 99%, respectively. The bioinformatics analysis showed that IGFBP-5 has an insulin growth factor-binding protein homologues (IB) domain and a thyroglobulin type-1 (TY) domain, four protein kinase C phosphorylation sites, five casein kinase II phosphorylation sites, three prenyl group binding sites (CaaX box). The IGFBP-5 gene was expressed in all the tested tissues including testis, brain, liver, lung, mammary gland, spleen, and kidney, suggesting that IGFBP-5 plays an important role in goat cells.

Phylogenetic Analysis of Agaricus blazei and Related Taxa by Comparing the Sequences of Internal Transcribed Spacers and 5.8S rDNA (Internal Transcribed Spacer와 5.8S ribosomal DNA의 염기서열 분석에 의한 Agaricus blazei와 근연종에 대한 계통분류학적인 연구)

  • 김기영;하명규;이태호;이재동
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.180-184
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    • 1999
  • Molecular spslemaucs of Agaricus species was investigated on the base of the sequences of the internal transcribed spaceriITS) regions in ribosomal DNA (rDNA). The sequences of the ITS region in 5 species and two group of Agaricus genus were resolved. In the phylogenetic trees. the species generally divided inlo two subclusters, refered to here as the group I and group 11. The group I consisted of Agaricus blazei ATCC 76739, Agarictrs blazei species cultivated in Korean hmings. Ago/-icus anmensis IMSNU 32049 and Agaricus can~pestris VPI-OKM 25665. Between Agaricus blazei NCC 76739 and the Agaricus blazei species cultivated in Korean farmings had the variation in lhe 5 nucleotide on the ITS regions. These varieties were presumed the variation by the geographic and cultivated conditions. In addition the subgroup of group I was formed by Agaricus arvensis LMSNU 32049 and Agaricus carnpests VPI-OKM 25665. The group IT included Agnrictrs bispoms CH 3004 and Agaricus pocillotor DUKE-J 173.

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