• Title/Summary/Keyword: S1 gene

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Nucleotide Sequence of the Putative Gene Encoding 30S Ribosomal Protein S1 from Brevibacterium ammoniagenes (Brevibacterium ammoniagenes의 30S 리보좀 단백질 S1을 코드하는 유전자의 염기서열)

  • 윤기홍;이미성;오영필;최정호
    • Microbiology and Biotechnology Letters
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    • v.28 no.3
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    • pp.147-151
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    • 2000
  • School of Food Biotechnology, W0050ng University, San 7-6, Jayang~dong. Dong-ku1 Taejon 300-100, Korea - The nucleotide sequence of approximately 2.4 kb immediately adjacent to ptsG gene coding for the glucose permease of Brevibacterium ammoniagenes was detennined. A putative open reading frame (ORP) of 1.467 nucleotides encoding a polypeptide of 489 amino acid residues and a TAA stop codon was identified. The deduced amino acid sequence of the ORF product has a high homology with the 30S ribosomal protein S 1 of Mycohacteriwn tuberculosis (83 % ). M leprae (74%), Streptomyces coelicola (77%), and Escherichia coli (40%). suggesting that the predicted product of ORF is a ribosomal protein S 1. The ORF is located at a distance of 266 nucleotides upstream from ptsC gene with a same translational direction.

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Liposome-Mediated Electric Gene Delivery into Fetal and Adult Gonads (Liposome을 매개로 한 태아 및 웅성 생식선으로의 전기적 유전자 도입)

  • Choi, S. C.;S. K. Choi;S. S. Choi;S. U. Kim;N. N. Cho;J. Y. Jung;C. S. Park;S. H. Lee;S. H. Lee
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.71-76
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    • 2004
  • Gene delivery is one of the keen interests in animal industry as well as research on gene functions. Some of the in vivo gene delivery techniques have been successively used in various tissues for the gene therapy and transgenesis. Despite intensive efforts, it still remains to overcome problems of limited local and regional administration and low transgene expression. To improve the efficiency of gene delivery, a new procedure was tested. We injected exogenous DNA containing LacZ into the female or male gonads and then pulsed electric field. Electroporated gonads showed positive X-gal staining in many seminiferous tubules of the porcine fetal gonads. Exogenously introduced LacZ genes were also expressed in female porcine gonad. In addition, we demonstrated efficient gene delivery in gonad of adult mouse. Furthermore, we succeed to generate genetically modified germline cells showing GFP and positive X-gal signals. The results suggest that the newly developed gene delivery is an effective way of in vivo transfection in mammals. The developed gene delivery procedure should be useful in producing transgenic animals when combined with primary cell culture and nuclear transplantation.

A case of pseudohypoaldosteronism type 1 with a mutation in the mineralocorticoid receptor gene

  • Lee, Se-Eun;Jung, Yun-Hye;Han, Kyoung-Hee;Lee, Hyun-Kyung;Kang, Hee-Gyung;Ha, Il-Soo;Choi, Yong;Cheong, Hae-Il
    • Clinical and Experimental Pediatrics
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    • v.54 no.2
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    • pp.90-93
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    • 2011
  • Pseudohypoaldosteronism type 1 (PHA1) is a rare form of mineralocorticoid resistance characterized in newborns by salt wasting with dehydration, hyperkalemia and failure to thrive. This disease is heterogeneous in etiology and includes autosomal dominant PHA1 owing to mutations of the NR3C2 gene encoding the mineralocorticoid receptor, autosomal recessive PHA1 due to mutations of the epithelial sodium channel (ENaC) gene, and secondary PHA1 associated with urinary tract diseases. Amongst these diseases, autosomal dominant PHA1 shows has manifestations restricted to renal tubules including a mild salt loss during infancy and that shows a gradual improvement with advancing age. Here, we report a neonatal case of PHA1 with a NR3C2 gene mutation (a heterozygous c.2146_2147insG in exon 5), in which the patient showed failure to thrive, hyponatremia, hyperkalemia, and elevated plasma renin and aldosterone levels. This is the first case of pseudohypoaldosteronism type 1 confirmed by genetic analysis in Korea.

Targeted Gene Disruption and Functional Complementation of Cytochrome P450 Hydroyxlase Involved in Cyclosporin A Hydroxylation in Sebekia benihana

  • Lee, Mi-Jin;Han, Kyu-Boem;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.14-19
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    • 2011
  • A cyclic undecapeptide-family natural product, cyclosporin A (CyA), which is one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex in a filamentous fungal strain named Tolypocladium niveum. Previously, structural modifications of cyclosporins such as a regionspecific hydroxylation at the $4^{th}$ N-methyl leucine in a rare actinomycetes called Sebekia benihana were reported to lead to dramatic changes in their bioactive spectra. However, the reason behind this change could not be determined since a system to genetically manipulate S. benihana has not yet been developed. To address this limitation, in this study, we utilized the most commonly practiced gene manipulation techniques including conjugation-based foreign gene transfer-and-expression as well as targeted gene disruption to genetically manipulate S. benihana. Using these optimized genetic manipulation systems, a putative cytochrome P450 hydroxylase (CYP) gene named CYP506, which is involved in CyA hydroxylation in S. benihana, was specifically disrupted and genetically complemented. The S. benihana${\Delta}$CYP506 exhibited a significantly reduced CyA hydroxylation yield as well as considerable yield restoration by functional complementation of the S. benihana CYP506 gene, suggesting that the genetically manipulated S. benihana CYP mutant strains may serve as a more efficient bioconversion host for various valuable metabolites including CyA.

Growth signaling and longevity in mouse models

  • Kim, Seung-Soo;Lee, Cheol-Koo
    • BMB Reports
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    • v.52 no.1
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    • pp.70-85
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    • 2019
  • Reduction of insulin/insulin-like growth factor 1 (IGF1) signaling (IIS) extends the lifespan of various species. So far, several longevity mouse models have been developed containing mutations related to growth signaling deficiency by targeting growth hormone (GH), IGF1, IGF1 receptor, insulin receptor, and insulin receptor substrate. In addition, p70 ribosomal protein S6 kinase 1 (S6K1) knockout leads to lifespan extension. S6K1 encodes an important kinase in the regulation of cell growth. S6K1 is regulated by mechanistic target of rapamycin (mTOR) complex 1. The v-myc myelocytomatosis viral oncogene homolog (MYC)-deficient mice also exhibits a longevity phenotype. The gene expression profiles of these mice models have been measured to identify their longevity mechanisms. Here, we summarize our knowledge of long-lived mouse models related to growth and discuss phenotypic characteristics, including organ-specific gene expression patterns.

Expression of Bacillus thringiensis HD-1 gene in rhizobacteria Pseudomonas fluorescens KR164 (근권 길항세균 Pseudomonas fluorescens KR164에 Bacillus thuringiensis HD-1 유전자의 삽입과 발현)

  • Kim, Yeong-Yil;Rhee, Young-Hwan;Kang, Heun-Soo
    • Applied Biological Chemistry
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    • v.35 no.4
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    • pp.227-231
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    • 1992
  • The plasmids pSUPBT and pSUPBTR were constructed with a vector pSUP2021 and the BT toxin gene in the plasmid pES 1. The plasmids constructed were introduced into the antagonistic rhizobacteria P. fluorescens KR164 by conjugation and P. fluorescens having pSUPBT and pSUPBTR were named P. fluorescens KR164(pSUPBT)#2, KR164(pSUPBT)#3, KR164(pSUPBTR)#2 and KR164(pSUPBTR)#3, respectively. The BT toxin gene were identified in all transformants by Southern hybridization and the final product of BT toxin gene was identified only in P. fluorescens KR164(pSUPBTR)#3 by SDS-PAGE. This crystal toxin protein were also observed in electron microscopy.

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Molecular Cloning and Characterization of Bovine HMGA1 Gene

  • Yu, S.L.;Chung, H.J.;Sang, B.C.;Bhuiyan, M.S.A.;Yoon, D.;Kim, K.S.;Jeon, J.T.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.11
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    • pp.1662-1669
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    • 2007
  • The high mobility group AT-hook1 (HMGA1) proteins are known to be related to the regulation of gene transcription, replication and promotion of metastatic progression in cancer cells. The loss of expression by disrupting the HMGA1 gene affects insulin signaling and causes diabetes in the mouse. Previously identified single nucleotide polymorphism (SNP) of HMGA1 was significantly associated with fat deposition traits in the pig. In this study, we identified 3,935 bp nucleotide sequences from exon 5 to exon 8 of the bovine HMGA1 gene and its mRNA expression was observed by quantitative real-time PCR. Six single nucleotide polymorphisms in the bovine HMGA1 gene were detected and the allele frequencies of these SNPs were investigated using the PCR-RFLP method in nine cattle breeds including Limousin, Simmental, Brown Swiss, Hereford, Angus, Charolais, Hanwoo, Brahman and Red Chittagong cattle. The map location showed that the bovine HMGA1 gene was also closely located with a previously identified meat quality QTL region indicating this gene is the most likely positional candidate for meat quality traits in cattle.

Enhanced Expression of Foreign Gene in Baculovirus-Infected Insect Cells Using a Silkworm Hemolymph (누에 체액을 이용한 외래 유전자의 발현효율 증대)

  • 우수동;김혜성
    • Journal of Sericultural and Entomological Science
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    • v.37 no.2
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    • pp.181-185
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    • 1995
  • To enhance expression of foreign gene by the novel expression vector, pBmKSK1, of Bombyx mori nuclear polyhedrosis virus, E. coli $\beta$-galactosidase gene expressing recombinant virus was infected in BmN-4 cells and various concentrations of silkworm hemolymph were added to the recombinant virus-infected BmN-4 cells containing fetal bovine serum. The expression efficiency of foreign gene was determined by $\beta$-galactosidase activity in the culture media. The results showed that the silkworm hemolymph was effective to expression of foreign gene in the BmN-4 cells, suggesting that the silkworm hemolymph could be substituted for fetal bovine serum in the BmN-4 cells to enhance expression of foreign gene.

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Functional Expression of the Internal Rotenone-Insensitive NADH-Quinone Oxidoreductase (NDI1) Gene of Saccharomyces cerevisiae in Human HeLa Cells

  • Seo, Byoung-Boo
    • Journal of Embryo Transfer
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    • v.25 no.1
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    • pp.35-42
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    • 2010
  • Many studies propose that dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I) is associated with neurodegenerative disorders, such as Parkinson's disease and Huntington's disease. Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. With a recombinant adeno-associated virus vector carrying the NDI1 gene (rAAV-NDI1) as the gene delivery method, we were able to attain high transduction efficiencies even in the human epithelial cervical cancer cells that are difficult to transfect by lipofection or calcium phosphate precipitation methods. Using a rAAV-NDI1, we demonstrated that the Ndi1 enzyme is successfully expressed in HeLa cells. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced HeLa cells were not affected by rotenone which is inhibitor of complex I, but was inhibited by flavone and antimycin A. The NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. In particular, in the NDI1-transduced cells, the yeast enzyme becomes integrated into the human respiratory chain. It is concluded that the NDI1 gene provides a potentially useful tool for gene therapy of mitochondrial diseases caused by complex I deficiency.