• Title/Summary/Keyword: early gene

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Effect of Cyclic GMP on Human Cytomegalovirus Gene Expression (Human Cytomegalovirus 유전자 발현에 Cyclic GMP의 영향)

  • Yoon, Joo-Hyun;Lee, Gyu-Cheol;Song, Byung-Hak;Kim, Young-Jin;Lee, Chan-Hee
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.261-269
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    • 1999
  • The relationship between second messenger cGMP and human cytomegalovirus (HCMV) replication was investigated. First, the intracellular level of cGMP ([cGMP]i) in HCMV-infected cells was measured. The [cGMP]i increased at early times after HCMV infection, reached maximum level at 12 hr and returned to basal level at 24 hr after virus infection, while [cGMP]i in mock-infected cells remained relatively unchanged. Increasing [cGMP]i resulted in enhanced transcription of HCMV major immediate early gene. For early gene expression, cGMP had varying effect. Expression of 1.2 kb RNA decreased and 2.2 kb RNA increased with increasing cGMP, while 2.7 kb RNA gene expression was not affected. HCMV early genes are regulated by immediate early gene, and the effect of cGMP on the regulatory effect of major immediate early gene on early genes was investigated. In the absence of cGMP, major immediate early gene repressed 2.7 kb RNA gene expression, while 1.2 kb RNA and 2.2 kb RNA early genes were not significantly affected. In the presence of $1\;{\mu}M$ cGMP, however, major immediate early gene stimulated the expression of three early genes.

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Expression of Human Cytomegalovirus Immediate Early US3 Gene in Human Fibroblast Cells

  • Lee, Gyu-Cheol;Lee, Chong-Kyo;Ahn, Jin-Hyun;Lee, Chan-Hee
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.24-30
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    • 2000
  • US3 gene is a member of the human cytomegalovirus (HCMV) immediate early gene. Although the precise functions of the US3 gene in HCMV replication and pathogenesis are not known, it has been reported to play a role in inhibiting major histocompatibility class I antigen presentation. For further knowledge of US3 gene expression, rabbit polyclonal antiserum of the US3 gene product was used for indirect immunofluorescence assay. In permissive human foreskin fibroblast (HFF) cells, US3 gene expression was detectable as crescent or half-moon shape in the perinuclear region at immediate early times after virus infection. HFF cells infected with mutant HCMV lacking US3 open reading frames were negative for US3 immunofluorescence assay. Double immunofluorescence assay using monoclonal antibody to gamma adaptin (specific for the Golgi complex) and rabbit anti-US3 antiserum revealed that US3 gene product could be localized to the Golgi complex. At later time after HCMV infection, US3 gene products were detected as globular aggregates in the cytosol. These aggregates were positive for gamma adaptin and stained with preimmune serum, suggesting a nonspecific reaction to the Golgi complex. Northern blot analysis revealed that transcription of US3 was observed only during immediate early times after virus infection (until 6 h postinfection). Therefore US3 gene expression appears to be confined to immediate early time and its gene products are localized to the Golgi complex as crescent shaped forms in the perinuclear cytoplasm.

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Genetic Engineering for Detection of Endocrine Disruption using I-18 C Gene Expression in Chironomus riparius

  • Kwak Inn-Sil
    • Korean Journal of Environmental Biology
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    • v.23 no.3 s.59
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    • pp.269-274
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    • 2005
  • The 2D/E gel analysis for polypeptide expression reflecting I-18 C gene (early-ecdysterone inducible gene) has conducted the emerged C. riparius adults from larval phase exposure to tebufenozide acting as an ecdysteroidal molting hormone. Control group, the amount of ORE II of the I-18 C gene was larger than that of ORE I of this gene. After treatments, ORE I of the I-18 C gene was overexpressed as the polypeptide, whereas ORF II of this gene was expressed as the polypeptide and was clearly reduced expression. Accordingly, we consider that tebufenozide exhibited endocrine disruptions related processing of ecdysteroid receptor protein reflecting ORF II of I-18 C gene. Also, earlier emergence day was related overexpressed polypeptide reflecting ORE I of I-18 C gene. In this study result, tebufenozide induced changing of physiological condition, and then polypeptide expression reflecting early-ecdysterone inducible I-18 C gene was different between control group and exposure group.

Copper, Zinc-Superoxide Dismutase (Cu/Zn SOD) Gene During Embryogenesis of Bombyx mori: Molecular Cloning, Characterization and Expression

  • Hong, Sun-Mee;Kang, Seok-Woo;Goo, Tae-Won;Kim, Nam-Soon;Lee, Jin-Sung;Nho, Si-Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • v.13 no.1
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    • pp.23-30
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    • 2006
  • BmCu/Zn SOD was isolated from early embryo of Bombyx mori using microarray analysis. The BmCu/Zn SOD gene was observed during the early embryonic stage with the strongest signal found at the unfertilizaion, fertilization and blastoderm stages. The BmCu/Zn SOD gene encodes a protein of 154 amino acids with a calculated Mr of 15 kDa. The deduced amino acid sequence of BmCu/Zn SOD indicated that the residues that form on the Cu/Zn binding site are conserved and that the sequence is a 60% identity to that of M. domestica. In a phylogenetic tree, Bm SOD was also close to Drosophila SODs rather than other insect SODs. The BmCu/Zn SOD gene exists as a single copy in the genome. Transcripts of BmCu/Zn SOD cDNA were identified by northern blot analysis. The expression of the BmCu/Zn SOD gene was observed weakly in most of larvae, pre-pupae, pupae and adult tissues. Also, the BmCu/Zn SOD gene was observed in early embryonic stage. Although the roles of SODs remains to be further elucidated, the high expression of BmCu/Zn SOD gene at before 24 h post fertilization suggests that this gene is of general importance during early embryogenesis in the Bombyx mod.

Very Early-Onset Inflammatory Bowel Disease: A Challenging Field for Pediatric Gastroenterologists

  • Arai, Katsuhiro
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.23 no.5
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    • pp.411-422
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    • 2020
  • With the increasing number of children with inflammatory bowel disease (IBD), very early-onset IBD (VEO-IBD), defined as IBD that is diagnosed or that develops before 6 years of age, has become a field of innovation among pediatric gastroenterologists. Advances in genetic testing have enabled the diagnosis of IBD caused by gene mutations, also known as monogenic or Mendelian disorder-associated IBD (MD-IBD), with approximately 60 causative genes reported to date. The diagnosis of VEO-IBD requires endoscopic and histological evaluations. However, satisfactory small bowel imaging studies may not be feasible in this small population. Both genetic and immunological approaches are necessary for the diagnosis of MD-IBD, which can differ among countries according to the available resources. As a result of the use of targeted gene panels covered by the national health insurance and the nationwide research project investigating inborn errors of immunity, an efficient approach for the diagnosis of MD-IBD has been developed in Japan. Proper management of VEO-IBD by pediatric gastroenterologists constitutes a challenge. Some MD-IBDs can be curable by allogenic hematopoietic stem cell transplantation. With an understanding of the affected gene functions, targeted therapies are being developed. Social and psychological support systems for both children and their families should also be provided to improve their quality of life. Multidisciplinary team care would contribute to early diagnosis, proper therapeutic interventions, and improved quality of life in patients and their families.

In Vitro Transcription Analyses of Autographa californica Nuclear Polyhedrosis Virus Genes

  • Huh, Nam-Eung
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.183-190
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    • 1994
  • Cell-free extracts prepared from cultured insect cells, Spodoptera. frugiperda, were analyzed for activation of early gene transcription of an insect baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV). The template DNA used for in vitro transcription assays contained promoter sites for the baculovirus genes that have been classified as immediate early ($\alpha$) or early genes. These genes are located in the HindIII-K/Q region of the AcNPV genome. Nuclei isolated from the AcNPV-infected Spodoptera frugiperda cells were also used for in vitro transcription analysis by RNase-mapping the labeled RNA synthesized from in vitro run-on reaction in the isolated nuclei. The genes studied by this technique were p26 and pl0 genes which were classified as delayed early and late gene, respectively. We found that transcription of the genes from the HindIII-K region was accurately initiated and unique in the whole cell extract obtained from uninfected cells, although abundance of the in vitro transcripts was reverse to that of in vivo RNA. With isolated nuclei transcription of the p26 gene was inhibited by $\alpha$-amanitin suggesting that the p26 gene was transcribed by host RNA polymerase II. However, transcription of the pl0 gene in isolated nuclei was not inhibited by $\alpha$-amanitin, but rather stimulated by the inhibitor. We also found that the synthesis of $\alpha$-amanitin-resistant RNA polymerase was begun before 6 hr p.i., the time point at which the onset of viral DNA replication as well as the appearance of a-amanitin-resistant viral transcripts were detected. These studies give us strong evidence to support the previous data that early genes of AcNPV were transcribed by host RNA polymerease III, while transcription of late genes was mediated at least by a novel $\alpha$-amanitin-resistant RNA polymerase.

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Molocular Cloning and Sequence Analysis of the Immediate Early Viral Gene, IE1, from Bombyx mori Nuclear Polyhedrosis Virus K1

  • Park, Hye-Jin;Lee, Kwang-Sik;Je, Yeon-Ho;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.1
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    • pp.43-49
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    • 2001
  • We have cloned and characterized an immediate early-1 gene, iel, which is activated immediately upon entrance of the viral genome into the cell nucleus, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. This gene encodes a protein 584 amino acids with a predicted molecular weight of 67 kDa. The promoter and coding regions of BmNPV-K1 ie1 showed high homology with Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain. The BmNPV-K1 ie1 was different from amino acid sequence at 4 positions in BmNPV T3. The location of ie1 gene in the BmNPV-K1 genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were confirmed by Nerthern hybridization analysis.

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Expression of Rice Chitinase Gene in Genetically Engineered Tomato Confers Enhanced Resistance to Fusarium Wilt and Early Blight

  • Jabeen, Nyla;Chaudhary, Zubeda;Gulfraz, Muhammad;Rashid, Hamid;Mirza, Bushra
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.252-258
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    • 2015
  • This is the first study reporting the evaluation of transgenic lines of tomato harboring rice chitinase (RCG3) gene for resistance to two important fungal pathogens Fusarium oxysporum f. sp. lycopersici (Fol) causing fusarium wilt and Alternaria solani causing early blight (EB). In this study, three transgenic lines TL1, TL2 and TL3 of tomato Solanum lycopersicum Mill. cv. Riogrande genetically engineered with rice chitinase (RCG 3) gene and their R1 progeny was tested for resistance to Fol by root dip method and A. solani by detached leaf assay. All the R0 transgenic lines were highly resistant to these fungal pathogens compared to nontransgenic control plants. The pattern of segregation of three independent transformant for Fol and A. solani was also studied. Mendelian segregation was observed in transgenic lines 2 and 3 while it was not observed in transgenic line 1. It was concluded that introduction of chitinase gene in susceptible cultivar of tomato not only enhanced the resistance but was stably inherited in transgenic lines 2 and 3.

EARLY SCREENING OF EXPRESSION OF SV40 DRIVEN LACZ INTRODUCED INTO BOVINE EMBRYOS

  • Nakamura, A.;Okumura, J.;Muramatsu, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.5
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    • pp.449-454
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    • 1995
  • The present study was conducted to assess gene expression of bacterial lacZ driven by the SV40 promoter at early developmental stages of bovine embryos. The lacZ gene was linearized with BamHI digestion and introduced into the pronucleus by microinjection at 20 hrs after the commencement of in vitro fertilization. Intact bovine blastocysts were not stained with X-Gal, suggesting that there is no endogenous beta-galactosidase activity in these blastocysts. In contrast, the bovine blastocyst cells microinjected with the lacZ gene exerted a characteristic greenish-blue color originating from the bacterial beta-galactosidase activity, albeit at a low rate, i.e. 2.1% of the total fertilized oocytes injected. It was concluded, therefore, that the lacZ gene driven by the SV40 promoter could be used for an indirect screening method in which the presence of transgene is evaluated from the product of transgene expression.

Association of Angiotensin Converting Enzyme I/D and ${\alpha}$-actinin-3 R577X Genotypes with Growth Factors and Physical Fitness in Korean Children

  • Kim, Kijin;Ahn, Nayoung;Cheun, Wookwang;Hong, Changbae;Byun, Jayoung;Joo, Youngsik
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.2
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    • pp.131-139
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    • 2015
  • This study analyzed the differences in aerobic and anaerobic exercise ability and growth-related indicators, depending on the polymorphism of the ACE and the ACTN3 genes, to understand the genetic influence of exercise ability in the growth process of children. The subjects of the study consisted of elementary school students (n=856, age $10.32{\pm}0.07yr$). The anthropometric parameters, physical fitness and growth factors were compared among groups of the ACE I/D or the ACTN3 R577X polymorphisms. There were no significant differences between the anthropometric parameters, physical fitness and growth factors for the ACE gene ID or the ACTN3 gene R577X polymorphism. However, the DD type of ACE gene was highest in the side step test (p<0.05), and the DD type was significantly higher than the II+ID type (p<0.05) in the early bone age. The combined group of the ACE gene II+ID and the ACTN3 gene XX type significantly showed lower early bone age (p< 0.05). This study did not find any individual or compounding effects of the polymorphism in the ACE I/D or the ACTN3 R577X polymorphisms on the anthropometric parameters, physical fitness and growth factors of Korean children. However, the exercise experience and the DD type of the ACE gene may affect the early maturity of the bones.