• Title/Summary/Keyword: H gene

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감마선 처리에 의한 스프레이형 국화 화색변이체로부터 Flavonoid 3'-Hydroxylase(F3'H) 유전자의 분리 및 특성 구명 (Isolation and Characterization of a Novel Flavonoid 3'-Hydroxylase (F3'H) Gene from a Chrysanthemum (Dendranthema grandiflorum) and Its Gamma-ray Irradiated Mutants)

  • 정성진;이긍주;김진백;김동섭;김상훈;강시용
    • 원예과학기술지
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    • 제30권2호
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    • pp.162-170
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    • 2012
  • 스프레이 국화품종 'Argus'와 감마선 조사에 의해 화색변이가 일어난 돌연변이체의 꽃잎으로부터 안토시아닌 생합성 경로에서 중요한 역할을 담당하는 신규 $DgF3'H$의 전장 cDNA와 genomic DNA를 분리하였다. 전장 cDNA는 1,527bp(509 아미노산)의 ORF를 포함하고 있으며, 원품종 'Argus'와 화색변이체 사이의 염기서열 상동성은 97% 이상을 나타내었다. Genomic DNA의 크기는 야생형 'Argus'에서 3,831bp이었고, 3가지 화색 변이체에서는 3,828부터 3,838bp의 크기를 나타내었다. $DgF3'H$ 유전자는 세 개의 exon사이에 두개의 intron을 갖고 있는 구조이고, 3'과 5' UTR 부분을 제외한 intron의 크기는 야생형 'Argus'에서 2,157bp이지만 3가지 화색 변이체에서는 2,155부터 2,159bp의 크기를 갖고 있었다. 이것은 감마선 조사에 의해 intron 부분의 유전자가 결실 또는 삽입된 것으로 추정된다. Southern 분석 결과 국화의 genome 내에서는 복수의 F3'H 유전자를 갖는 것이 확인되었다. $DgF3'H$ 유전자의 발현 정도를 분석한 결과, 연분홍의 'Argus'와 두 개의 보라색 변이체(AM1 and AM3)에서 높게 발현되었으나 흰색 변이체(AM2)에서는 매우 약하게 발현되었으며, 염기서열 변이에 의한 F3'H 유전자의 구조적 차이가 화색의 변이에 관련된 것으로 추정되었다. 국화 'Argus' 및 화색 변이체를 이용하여 본 연구에서 분리한 신규 F3'H 유전자의 구조 및 유전자 발현 등을 포함하는 유전정보들은 화색 변이의 유전적 기작을 밝히는데 중요한 자료로 이용될 것으로 기대되나 향후 다른 유전적 발현요소들이 국화의 F3'H 유전자의 발현에 관여하는지에 관한 추가적인 연구가 필요하다고 하겠다.

Increase of Yeast Survival under Oxidative Stress by the Expression of the Laccase Gene from Coprinellus congregatus

  • Kim, Dong-Sik;Kwak, Eun-Jung;Choi, Hyoung-T.
    • Journal of Microbiology
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    • 제44권6호
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    • pp.617-621
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    • 2006
  • Coprinellus congregatus secreted a laccase isozyme when the culture was transferred to an acidic liquid medium (pH 4.1). The laccase cDNA gene (clac2) was used as a probe for cloning of the genomic laccase gene (lac2) including the promoter (Plac2). The open reading frame (ORF) of lac2 had 526 deduced amino acids and four conserved copper binding domains as other fungal laccases. Recombinant plasmid (pRSlac2p-cDNA) of lac2 cDNA with its own promoter was transformed in Saccharomyces cerevisiae. Expression of the transformed lac2 gene was induced by oxidative stress ($H_2O_2$) in yeast and the survival rate of the transformed yeast strain was greatly increased when compared with that of the control strain transformed with pRS316 yeast vector.

Effects of Homogentisic Acid and Natural Products Derived from Pinellia ternata on Secretion, Production and Gene Expression of MUC5AC Mucin from Cultured Airway Epithelial Cells

  • Lee, Hyun Jae;Lee, Choong Jae
    • Natural Product Sciences
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    • 제23권1호
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    • pp.29-34
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    • 2017
  • In this study, we investigated whether adenosine, adenine, uridine and homogentisic acid derived from Pinellia ternata affect the secretion, production and gene expression of MUC5AC mucin from airway epithelial cells. Confluent NCI-H292 cells were pretreated with adenosine, adenine, uridine or homogentisic acid for 30 min and then stimulated with PMA (phorbol 12-myristate 13-acetate) for 24 h. The MUC5AC mucin gene expression, mucin protein production and secretion were measured by RT-PCR and ELISA, respectively. The results were as follows: (1) Adenine and homogentisic acid decreased PMA-induced MUC5AC mucin gene expression, although adenosine and uridine did not affect the mucin gene expression; (2) Adenosine, adenine, uridine and homogentisic acid inhibited PMA-induced MUC5AC mucin production; (3) Homogentisic acid inhibited the secretion of MUC5AC mucin from NCI-H292 cells. These results suggest that, among the four compounds examined, homogentisic acid showed the regulatory effect on the steps of gene expression, production and secretion of mucin, by directly acting on airway epithelial cells.

Nucleotide Sequence on Upstream of the cdd Locus in Bacillus subtilis

  • JONG-GUK KIM;KIM, KYE-WON;SEON-KAP HWANG;JOO-WON SUH;BANG-HO SONG;SOON-DUCK HONG
    • Journal of Microbiology and Biotechnology
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    • 제5권3호
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    • pp.125-131
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    • 1995
  • A 3, 346 bp of the cdd upstream region in Bacillus subtilis was sequenced from the pSO1 (Song BH and J Neuhard. 1989. Mol. Gen. Genet 216: 462-468) and sequence homology was searched to the known genes in Genbank and European Molecular Biology Laboratory databanks. Five complete and one truncated putative coding sequences deduced from the nucleotide sequence were found through the ORF searching by Genetyx and Macvector software, and one of them was identified as the dgk (diacylglycerol kinase) gene and another, a truncated one, as the phoH (phosphate starvation-inducible gene) gene. The B. subtilis dgk gene, having a role for response to several environmental stress signals, revealed an open reading frame of 134 amino acids with 43.1% of sequence identity to the Streptococcus mutans dgk gene. The carboxy terminal 59 residues of the truncated phoH gene showed 52.7% and 34.5% of sequence identity in amino acids with the corresponding genes of Mycobacterium leprae and Escherichia coli. The four remaining coding sequences consisting of 115, 421, 91, and 91 residues were thought to be unknown ORFs because they have no significant similarity to known genes.

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Human Pluripotent Stem Cell-Derived Retinal Organoids: A Viable Platform for Investigating the Efficacy of Adeno-Associated Virus Gene Therapy

  • Hyeon-Jin Na;Jae-Eun Kwon;Seung-Hyun Kim;Jiwon Ahn;Ok-Seon Kwon;Kyung-Sook Chung
    • International Journal of Stem Cells
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    • 제17권2호
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    • pp.204-211
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    • 2024
  • With recent advances in adeno-associated virus (AAV)-based gene therapy, efficacy and toxicity screening have become essential for developing gene therapeutic drugs for retinal diseases. Retinal organoids from human pluripotent stem cells (hPSCs) offer a more accessible and reproducible human test platform for evaluating AAV-based gene therapy. In this study, hPSCs were differentiated into retinal organoids composed of various types of retinal cells. The transduction efficiencies of AAV2 and AAV8, which are widely used in clinical trials of inherited retinal diseases, were analyzed using retinal organoids. These results suggest that retinal organoids derived from hPSCs serve as suitable screening platforms owing to their diverse retinal cell types and similarity to the human retina. In summary, we propose an optimal stepwise protocol that includes the generation of retinal organoids and analysis of AAV transduction efficacy, providing a comprehensive approach for evaluating AAV-based gene therapy for retinal diseases.

Characterization and Expression in Escherichi coli of Streptococcus pneumoniae FtsH

  • Kim, Hee-Soo;Lee, Jae-Jung
    • 대한미생물학회지
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    • 제35권2호
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    • pp.109-115
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    • 2000
  • FtsH is a membrane-bound, ATP-dependent metalloprotease that is involved in a variety of cellular functions including the regulation of responses to heat and stress shock. Previously, we had cloned and sequenced pneumococcal ftsH gene whose deduced amino acid sequence was very similar to those of several gram-positive bacteria and Escherichia coli, except for the N-terminal domain that was responsible for membrane anchoring. In order to better understand the role of Streptococcus pneumoniae FtsH, we expressed pneumococcal ftsH gene in Escherichia coli. When it was expressed from a strong promoter, $P_{tac}$, a considerable amount of the recombinant FtsH was produced, although the prolonged induction resulted in not only accumulation of breakdown products but also ceasing of the further growth of E. coli host. This indicated that the expression of the exogenous ftsH gene was tightly regulated since the excessive FtsH appeared detrimental to bacterial cells. In Western blotting, the pneumococcal FtsH protein, whether native or recombinant, was reactive to anti-E. coli FtsH serum. The observation that FtsH proteins were well conserved throughout the bacterial kingdom and its expression level was fine-tuned suggests an important role for this protein in the stress adaptation which may be related to infecting process by pneumococci.

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Molecular Characterization and Expression Patterns of Porcine Eukaryotic Elongation Factor 1 A

  • Wang, H.L.;Wang, H.;Zhu, Z.M.;Yang, S.L.;Fen, S.T.;Li, Kui
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권7호
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    • pp.953-957
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    • 2006
  • The eukaryotic elongation factor 1 A (EEF1A) participates in protein synthesis by forming the eEF1A GTP tRNA complex to deliver aminoacyl-tRNA to the A site of ribosomes. This study described cDNA sequences and partial genomic structure of porcine EEF1A1. The porcine EEF1A1 gene encoded a protein with 462 amino acids, which shared complete homology with human, chimpanzee and dog. The temporal expression pattern showed the diversity of EEF1A1 level in mRNA was relatively minor in prenatal embryo skeletal muscle, however, the expression decreased during aging after birth in skeletal muscle of the Chinese Tongcheng pig. The spatial expression patterns indicated that the gene expressed in skeletal muscle, heart, lung, liver, kidney, fat and spleen. In addition, we assigned the gene to porcine chromosome 1 using a radiation hybrid panel.