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Zygotic Expression of c-myc Gene in Mouse Early Embryos: Functional Role of c-myc Promoter (생쥐 초기배아에서 c-myc Proto-Oncogene Promoter의 기능적 활성화)

  • Park, Ki-Soo;Kang, Hae-Mook;Shim, Chan-seob;Sun, Woong;Kim, Jae-man;Lee, Young-Ki;Kim, Kyung-jin
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.550-556
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    • 1995
  • The c-myc proto-oncogene is Involved In the control of normal cell proliferation and differentiation of many cell lineages. Although it has heen suggested that c-myc may play an important role in the mammalian early development, it Is unclear whether the embryonic c-myc mRNA is originated from zygotic gene expression or stored maternal message. Thus, we have construded expression vectors, In which the 5, flanking sequences including c-myc promoter region and a large non-coding exon I are fused 'sith E. coli lacZ gene that encedes $\beta$-galactosldase as a reporter. As c-myc exon I contains a modulatory sequence, we designed t, vo types of vectors (pcmyc.Gall and pcmyc-Ga12) to examine the role of exon I in c-myc expression. The former contains the complete exon I and the later has a deletion in 40 bp of modulator sequence located In the exon I of c-myc These vectors were microInjected into fertilized one-cell embryos and $\beta$-galactosidase activity was examined by X-gal staining during early embryogenesis. $\beta$-galactosidase activity derived from c-myc promoter was decreased at two-cell stage. The expression level directed by pcmyc- Ga12 was similar to that of pcmyc-Gal1, indicating that the medulatory sequence in exon I may not be Involved at least In the regulation of embryonic c-myc expression. In summary, the present study indicates that the c-myc promoter is functional at the early stage embryo, and the regulation of c-myc expression is under the control of "zygotic" clock of preimplantation mouse embryos.e embryos.

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Phylogenetic Analysis of 680 Prokaryotes by Gene Content (유전자 보유 계통수를 이용한 원핵생물 680종의 분석)

  • Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.26 no.6
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    • pp.711-720
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    • 2016
  • To determine the degree of common genes and the phylogenetic relationships among genome-sequenced 680 prokaryotes, the similarities among 4,631 clusters of orthologous groups of protein (COGs)’ presence/ absence and gene content trees were analyzed. The number of COGs was in the range of 103–2,199 (mean 1377.1) among 680 prokaryotes. Candidatus Nasuia deltocephalinicola str. NAS-ALF, an obligate symbiont with insects, showed the minimum COG, while Pseudomonas aeruginosa PAO1, an opportunistic pathogen, represented the maximum COG. The similarities between two prokaryotes were 49.30–99.78 % (mean 72.65%). Methanocaldococcus jannaschii DSM 2661 (hyperthermophilic and autotrophic, Euryarchaeota phylum) and Mesorhizobium loti MAFF303099 (mesophilic and symbiotic, alpha-Proteobacteria class) had the minimum amount of similarities. As gene content may represent the potential for an organism to adapt to each habitat, this may represent the history of prokaryotic evolution or the range of prokaryotic habitats at present on earth. COG content trees represented the following. First, two members of Chloroflexi phylum (Dehalogenimonas lykanthroporepellens BL-DC-9 and Dehalococcoides mccartyi 195) showed a greater relationship with Archaea than other Eubacteria. Second, members of the same phylum or class in the 16S rRNA gene were separated in the COG content tree. Finally, delta- and epsilon-Proteobacteria were in different lineages with other Proteobacteria classes in neighbor-joining (NJ) and maximum likelihood (ML) trees. The results of this study would be valuable to identifying the origins of organisms, functional relationships, and useful genes.

Construction of Transgenic Silkworms Expressing Human Stem Cell Factor (hSCF) (인간 유래 Stem Cell Factor (hSCF) 재조합단백질이 발현되는 누에형질전환체 제작)

  • Kim, Sung-Wan;Yun, Eun-Young;Kim, Seong-Ryul;Park, Seung-Won;Kang, Seok-Woo;Kwon, O-Yu;Goo, Tae-Won
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1726-1731
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    • 2011
  • Human Stem Cell Factor (hSCF) is a cytokine that binds to the c-Kit receptor and plays an important role in hematopoiesis, spermatogenesis, and melanogenesis. To produce the human Stem Cell Factor (hSCF) recombinant protein, we constructed a germline transgenic silkworm using the piggyback vector. The expression of the hSCF gene was driven by the Drosophila heat shock protein 70 (dHsp70) promoter. 3XP3 promotor-driven EGFP was used as a marker which allowed us to rapidly distinguish the transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,020 eggs of bivoltin silkworms, Keomokjam. We obtained approximately 22 G1 broods that were EGFP-positive. The expression of the hSCF gene in the transgenic silkworm was analyzed by SDS-PAGE and immunoblotting. Also, analysis of insertion sites into the silkworm genome using inverse PCR showed that exogenous DNA was inserted into the transgenic silkworm genome. These results show that successfully constructed transgenic silkworm expresses the hSCF recombinant protein.

Effect of hnRNP-like protein THO4 on growth and mRNA export in fission yeast (분열효모에서 hnRNP-유사 단백질인 THO4가 생장 및 mRNA 방출에 미치는 영향)

  • Park, Jin Hee;Lee, Sojeong;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.91-97
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    • 2018
  • The evolutionally conserved TREX complex member, Yra1/ALY, belongs to the REF (RNA and export factor binding proteins) family of hnRNP-like proteins, which has been implicated in multiple processes including transcription, nuclear RNA stability, and mRNA export. Fission yeast, Schizosaccharomyces pombe, genome encodes two members of REF proteins. In addition to Mlo3 known previously as an mRNA export factor, there is the other REF protein, Tho4, which is predicted as a component of THO complex. Here we showed that deletion of tho4 (SPBC106.12c) gene does not inhibit both growth and nuclear mRNA export. However, overexpression of tho4 displays growth retardation and slight accumulation of $poly(A)^+$ RNA in the nucleus. Neither ${\Delta}tho4$ ${\Delta}mlo3$ nor ${\Delta}tho4$ ${\Delta}mex67$ double mutants exhibit additive growth defect. Moreover, yeast two-hybrid and co-immunoprecipitation analysis did not show that the Tho4 protein interacted with any members of TREX complex and mRNA export factor Rae1. Contrary to expectation, these observations support that the S. pombe Tho4 is not a component of TREX complex, and not directly involved in bulk mRNA export from the nucleus.

An extension of multifactor dimensionality reduction method for detecting gene-gene interactions with the survival time (생존시간과 연관된 유전자 간의 교호작용에 관한 다중차원축소방법의 확장)

  • Oh, Jin Seok;Lee, Seung Yeoun
    • Journal of the Korean Data and Information Science Society
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    • v.25 no.5
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    • pp.1057-1067
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    • 2014
  • Many genetic variants have been identified to be associated with complex diseases such as hypertension, diabetes and cancers throughout genome-wide association studies (GWAS). However, there still exist a serious missing heritability problem since the proportion explained by genetic variants from GWAS is very weak less than 10~15%. Gene-gene interaction study may be helpful to explain the missing heritability because most of complex disease mechanisms are involved with more than one single SNP, which include multiple SNPs or gene-gene interactions. This paper focuses on gene-gene interactions with the survival phenotype by extending the multifactor dimensionality reduction (MDR) method to the accelerated failure time (AFT) model. The standardized residual from AFT model is used as a residual score for classifying multiple geno-types into high and low risk groups and algorithm of MDR is implemented. We call this method AFT-MDR and compares the power of AFT-MDR with those of Surv-MDR and Cox-MDR in simulation studies. Also a real data for leukemia Korean patients is analyzed. It was found that the power of AFT-MDR is greater than that of Surv-MDR and is comparable with that of Cox-MDR, but is very sensitive to the censoring fraction.

Cloning and Sequencing of the pelCl Gene Encoding Pectate Lyase of Erwinia carotovora subsp. carotovora LY34 (Erwinia carotovora subsp. carotovora LY34에서 pelCI 유전자 클로닝)

  • Lim, Sun-Tech;Park, Yong-Woo;Yun, Han-Dae
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.380-387
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    • 1997
  • Phytopathogenic Erwinia carotovora subsp. carotovora (Ecc) LY34 causes plant tissue maceration by secretion of pectinolytic enzymes such as pectate Iyase (PL) existed as multiple isoenzyme form. Genomic DNA from Ecc LY34 was digested with Sau3Al and ligated into the BamHI site of pBluescript ll $SK^+$. Among them, a clone hydrolyzing polypectate was selected and its DNA was digested with BamHI. Through the subsequent subcloning the resulting 3.1 kb fragment, corresponding to a peICI, was subcloned into pLYPA 100. The structural organization of a peICI gene encoding a 374 amino acid residues consists of an open reading frame (ORF) of 1,122 bp commencing with a ATG start codon and followed by a TAA stop codon. PeICI contained a typical prokaryotic signal peptide of 22-amino acid. Since the deduced amino acid sequences of PeICl protein was very similar to those of PelIII of Erwinia carotovora subsp. carotovora, and to those of Pel3 of Erwinia carotovora subsp. atroseptica, and to those of PeIC of Erwinia carotovora subsp. carotovora, it belong to the same family PLbc group. The 374-amino acld PeICI had a calculated Mr of 40,507 and pI of 7.60.

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Characterization of the Neurospora crassa rcm-1 Mutants (Neurospora crassa rcm-1 돌연변이체의 특성)

  • Kim Sang-Rae;Lee Bheong-Uk
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.246-254
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    • 2005
  • Analysis of the complete genome of Neurospora crassa reveals that at least 19 proteins contain tetratricopeptide repeat (TPR) motifs. One of them shows over $60\%$ homology to Ssn6 of Saccharomyces cerevisiae, a universal repressor that mediates repression of genes involved in various cellular processes. Mutant strains generated by RIP (repeat-induced point mutation) process showed four distinctive vegetative growth patterns and slow growth in various rates. Firstly, a mutant showed denser mycelial growth, yellow, csp, and looked like ropy mutant. Secondly, slower growth, dense mycelial, and conidial phenotype. Thirdly, extremely slower growth and aconidial. And finally, flat, tittle aerial hyphae, acon, and similar with a rco-1 RIP mutant. They are all male-fertile, yet female-sterile and produced little or no perithecium. It seems that various phenotypes were occurred depending upon mostly likely, the degree of RIP. These results indicate that this gene may be involved in several cellular possess during vegetative growth, and asexual and sexual development. Therefore it is pleiotropic. Sequence analysis of cDNA shows that it encodes a putative 102 kDa protein composed of 917 amino acids, and has six introns. It is designated rcm-1 (regulation of conidiation and morphology).

Replication and Pathogenesis of Plaque Morphology Mutants Derived from Vero Cells with Japanese Encephalitis Virus Persistency (지속감염세포에서 분리된 일본뇌염바이러스 Plaque Morphology Mutants의 복제 및 감염특성)

  • 윤성욱;정용석
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.221-229
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    • 2002
  • Japanese encephalitis virus (JEV) persistence was established and maintained in Vero cell culture for over 1 year. Eleven clones of plaque morphology mutant JEV, with large and small plaque sizes, were obtained from the cell culture supernatant. Genomic RNA replication efficiency of the mutants in naive Vero cell appeared to correspond to their different plaque sizes. No significant changes in envelop protein ORF or in non-coding regions at both ends of the RNA genome suggested that there could be an unidentified factor(s) playing role in JEV attenuation. Unlike to the replication of wild-type JEV, the mutants did not induce severe degree of cytopathic effect in Vero cells upon infection. While obvious decrease of Bcl-2 and its mRNA expression and sharp increase of p53 in naive Vero cells infected with either wild-type JEV or the large plaque-forming mutant, those changes were not observed with the small plaque-forming one. Together with these observation, internucleosomal DNA fragmentation and chromosomal DNA profile in the Vero cells infected with the mutants suggest that an overall changes in cytopathic effect in the plaque morphology mutants-infected cells should be primarily due to the reduced genomic RNA replication and the compromised degree of p53-independent apoptosis by the virus infection at least in part.

An Increment of Crossing Efficiency with Consideration of Pollen Viability Analysis in Rose (장미 교배 효율 증대를 위한 화분 임성 검정)

  • Hwang, Yoon Jung;Song, Chang Min;Kwon, Min Kyung;Kim, Sung Tae;Kim, Won Hee;Han, Youn Yol;Han, Tae Ho;Lim, Ki Byung
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.3
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    • pp.193-200
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    • 2010
  • Current rose cultivars are all composed of heterozygous genome due to long history of out crossing including interspecific hybridization. It has been adapted by artificial selection and crossing by breeders that mainly based on the crossing with fertile pollen derived from inter- or intra-specific hybridization. Pollen viability and germination ability tests provide valuable information for the designing of parentage for more successful breeding efficacy. In this study, we tested the pollen viability and germination ability in seven rose cultivars to find any relationship among several factors including pollen size, ploidy levels, and crossing compatibility. The pollen viability showed wide ranges from 39% 'Pinocchio' as minimum to 82% 'Scarlet Mimi' as maximum, whereas pollen germination rate were from 1% 'Mini Rosa' to 41% 'Scarlet Mimi' as a highest. Pollen size ranged from 41.3 to $45.4{\mu}m$ in large sized pollen and 30.7 to $37.4{\mu}m$ in small sized pollen. The mean diameter of large sized pollen is approximately 10-40% bigger than that of small sized pollen. There are positive relationships among ploidy level, total chromosome length, and pollen size. Crossing list showed that seed setting ratio and seed germination were related to pollen viability, pollen germination, and ploidy level.

Development of Salt-Tolerant Transgenic Rice Using Soybean PR10 Gene (콩의 Pathogenesis-Related 10 유전자를 이용한 내염성 벼 형질전환 계통 개발)

  • Kim, Hyo Jin;Baek, So Hyeon;Shin, Woon Chul;Seo, Chun Sun;Park, Myoung Ryoul;Ko, Jae Kwon;Yun, Song Joong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.540-546
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    • 2010
  • This study was conducted to understand the role of soybean pathogenesis-related 10 (GmPR10) gene in salt tolerance and to develop salt-tolerant rice using GmPR10 cDNA. GmPR10 transgene was expressed constitutively in the shoot and root of the $T_1$ transgenic rice plants. Interestingly, however, the levels of the transgene expression were increased temporally up to over four- to five-fold in the shoot and root by 125 mM NaCl treatment, peaking at six hours after the treatment and decreasing thereafter. Electrolyte leakage of leaf cells under 125 mM NaCl treatment was lower in all the transgenic lines than in the control variety, Dongjin-byeo. Ability of seedlings to recover from 125 mM NaCl treatment for two weeks was higher in the transgenic plants than in the control plants. These results demonstrated that GmPR10 had function to increase cell integrity and promote growth under the saline stress imposed by NaCl. The transgenic line GmPR10-3 which showed highest ability to recover from the saline stress could be used as a potential source for salt tolerance in rice breeding programs.