• Title/Summary/Keyword: Modifying gene

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Production of the taste-modifying protein, miraculin, in transgenic lettuce

  • Ezura, Hiroshi;Sun, Heyon-Jin
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.126-131
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    • 2005
  • Richadella dulcifica, a native shrub in tropical West Africa, gives red berries that have the unusual property of modifying a sour taste into a sweet taste. The red berries contain a taste-modifying protein named miraculin. A synthetic gene encoding miraculin was placed under the control of constitutive promoters and transferred to lettuce. High expression of miraculin was obtained, with accumulation of up to 1% total soluble protein in lettuce leaf. In addition, the miraculin expressed in lettuce possesses a taste-modifying activity.

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Novel Therapeutic Approaches to Mucopolysaccharidosis Type III

  • Yang, Aram
    • Journal of mucopolysaccharidosis and rare diseases
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    • v.5 no.1
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    • pp.22-28
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    • 2021
  • Mucopolysaccharidosis type III (MPS III) or Sanfilippo disease is an orphan-inherited lysosomal storage disease. It is one of the most common MPS subtypes. The classical presentation is an infantile-onset neurodegenerative disease characterized by intellectual regression, behavioral and sleep disturbances, loss of ambulation, and early death. Unlike other MPS, no disease-modifying therapy has been approved. Here, we review the curative therapy developed for MPS III, from historically ineffective hematopoietic stem cell transplantation and substrate reduction therapy to the promising enzyme replacement therapy or adeno-associated/lentiviral vector-mediated gene therapy. Preclinical studies are presented with recent translational first-in-man trials. We also present experimental research with preclinical mRNA and gene-editing strategies. Lessons from animal studies and clinical trials have highlighted the importance of early therapy before extensive neuronal loss. Disease-modifying therapy for MPS III will likely mandate the development of new early diagnosis strategies.

The Dopamine $D_2$ Receptor Locus as a Modifying Gene in Korean Schizophrenia, Alcoholism and Drug Addiction (정신분열증, 알코올중독, 약물중독에서 도파민 $D_2$ 수용체 유전자의 조절유전자(modifying gene)로서의 역할 - 충동적.강박적.탐닉적 행동을 나타내는 정신질환들에서 도파민 $D_2$ 수용체의 조절유전자로서의 역할 -)

  • Jung, Hyun-Mo;Lee, Hong-Seock;Chang, Dong-Won;Lee, Min-Soo
    • Korean Journal of Biological Psychiatry
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    • v.4 no.2
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    • pp.225-233
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    • 1997
  • The authors attempted to examine the allelic association between the A1 allele of Dopamine $D_2$ receptor and schizophrenia, alcoholism, drug addiction in Koreans. Schizophrenic patients(n=31), alcoholism(n=65), drug addiction(n=18) and controls(n=52) were examined by case-control study for distribution of the TaqI polymorphism of the dopamine $D_2$ receptor gene in Korean population to minimize the effect of racial differencies in gene frequencies. In schizophrenics, the numbers of schizophrenics with A1A1, A1A2, A2A2 were 9(29.0%), 15(48.4%) and 7(22.6%) respectively and in alcoholics with A1A1, A1A2, A2A2 were 14(21.5%), 36(55.4%) and 15(23.1%) respectively and in drug addiction with A1A1, A1A2, A2A2 were 2(11.1%), 10(55.6%) and 6(33.3%) respectively and in controls with A1A1, A1A2, A2A2 were 4(7.6%), 24(46.2%) and 24(46.2%) respectively. The prevalence of the A1 allele in schizophrenics, alcoholics, drug addiction and controls were 77%, 76.9%, 67% and 53.8% respectively. And the frequency of the A1 allele in schizophrenics, alcoholics, drug addiction and controls were 0.53, 0.49 0.39 and 0.31 respectively. There was significant difference in the frequency of the A1 allele between schizophrenics, alcoholics and controls. We also classified our alcoholic population. For classification by severity, we used the median MAST score 30 in our samples. There was also significant difference in the frequency of the A1 allele between less severe group(0.42) and more severe group(0.57). This data suggest that the A1 allele is associated with schizophrenia and alcoholism in Koreans. Furthermore the prevalence of the A1 allele increassed in more severely affected alcoholics. The authors conclude that our data support an allelic association between the A1 allele at dopamine $D_2$ receptor and schizophrenia, alcoholism. These results suggest the A1 allele of the $DRD_2$ gene is associated with a number of behavior disorders in which it may act as a modifying gene rather than as the primary etiological agent.

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Application of sweet and taste modifying genes for development in plants: current status and prospects

  • Akter, Shahina;Huq, Md. Amdadul;Jung, Yu-Jin;Cho, Yong-Gu;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.43 no.4
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    • pp.397-404
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    • 2016
  • Sweet and taste modifying proteins are natural alternatives to synthetic sweeteners and flavor enhancers, and have been used for centuries in different countries. Use of these proteins is limited due to less stability and availability. However, recent advances in biotechnology have enhanced their availability. These include production of sweet and taste modifying proteins in transgenic organisms, and protein engineering to improve their stability. Their increased availability in the food, beverage or medicinal industries as sweeteners and flavor enhancers will reduce the dependence on artificial alternatives. Production of transgenic plants using sweet and taste modifying genes, is an interesting alternative to the extraction of these products from natural source. In this review paper, we briefly describe various sweet and taste modifying proteins (such as thaumatin, monellin, brazzein, curculin and miraculin), their properties, and their application for plant development using biotechnological approaches.

Spreading of Aminoglycoside-Modifying Enzymes among Escherichia coli Isolated from Clinical Specimens in Chungcheong Province (충청지역의 임상검체로부터 분리된 대장균에 Aminoglycoside-Modifying Enzymes 확산)

  • Sung, Ji Youn;Kwon, Pil Seung
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.2
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    • pp.136-142
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    • 2020
  • The antimicrobial resistance rate in bacteria has increased over the last several decades. The transfer of antimicrobial resistant determinants on mobile genetic elements could cause the accelerated emergence and spread of multidrug resistant bacteria. This study investigated the aminoglycoside resistance determinants transferred by mobile genetic elements in a total of 33 aminoglycoside non-susceptible E. coli isolated from clinical specimens in Chungcheong province. 16S ribosomal RNA methyl-transferases (RMTases) and aminoglycoside-modifying enzyme (AME) genes were detected via PCR and DNA sequencing. The most common AME genes were aac(3')-II gene (54.5%), followed by aph(3')-Ia (18.2%) and aac(6')-Ib (15.2%). None of the evaluated RMTase genes were detected in the 33 isolates. Seventeen of the 18 isolates harboring aac(3')-II gene were resistant to gentamicin, and 16 of them were resistant to tobramycin. The 5 isolates harboring aac(6')-Ib gene were all resistant to tobramycin. In this study, we confirmed that one of the important mechanisms of aminoglycoside resistance in E. coli isolated from human is the acquisition of AME genes. Continuing investigations of antimicrobial resistant determinants in bacteria isolated from human may be required to prevent dissemination of antimicrobial resistant bacteria.

Overproduction and Secretion of $\beta$-Glucosidase in Bacillus subtilis

  • Kim, Jeong-Hyun;Lee, Baek-Rak;Moo, young-Pack
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.141-145
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    • 1998
  • Overproduction of intracellular ${\beta}$-glucosidase was attempted by modifying the promoter region of a ${\beta}$-glucosidase gene cloned from Cellulomonas fimi and expressing it in Bacillus subtilis DB 104. A strong engineered promoter, BJ27UΔ88, was fused to the ${\beta}$-glucosidase gene after removing its native promoter. An effective Shine-Dalgamo sequence (genel0 of phage T7) was inserted between the promoter and the ${\beta}$-glucosidase structural gene. The modified gene was overexpressed in B. subtilis and produced 1121.5 units of ${\beta}$-glucosidase per mg protein which is about $12\%$ of total intracellular protein. Secretion of overproduced intracellular ${\beta}$-glucosidase was attempted by using the signal sequence of the Bacillus endoglucanase gene as well as an in-frame hybrid protein of endoglucanase. The hybrid protein was normally secreted into the culture medium and still retained ${\beta}$-glucosidase activity.

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Expression of the Gene Encoding Firefly Luciferase Using Bombyx mori Nucleopolyhedrovirus Vector

  • Woo, Soo-Dong;Cho, Kook-Ho;Jin, Byung-Rae;Boo, Kyung-Saeng;Kang, Seok-Kwon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.1 no.1
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    • pp.53-58
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    • 2000
  • A cDNA encoding the luciferase of firefly Luciola lateralis was cloned downstream from the polyhedrin gene promoter of Bombyx mori nucleopolyhedrovirus and expressed in B. mori cells (BmN-4). The coding soquence for luciferase was inserted into pBmKSK2 rectors) which was reconstructed from the polyhedrin-based transfer vector pBmKSKl by modifying cloning sites. Recombinant virus, BmK2-LUCDF, containing the luciferase gene was selected and purified in BmN-4 cells. The emission of luminescence by luciferase was only detected in BmK2-LUCDF-infected cell extracts. This result indicates that the cloned new luciferase gene of firefly L. lateralis can be expressed efficiently in baculovirus expression system and used as a useful reporter gene.

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A Mutation of a Putative NDP-Sugar Epimerase Gene in Ralstonia pseudosolanacearum Attenuates Exopolysaccharide Production and Bacterial Virulence in Tomato Plant

  • Hyoung Ju Lee;Sang-Moo Lee;Minseo Choi;Joo Hwan Kwon;Seon-Woo Lee
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.417-429
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    • 2023
  • Ralstonia solanacearum species complex (RSSC) is a soil borne plant pathogen causing bacterial wilt on various important crops, including Solanaceae plants. The bacterial pathogens within the RSSC produce exopolysaccharide (EPS), a highly complicated nitrogencontaining heteropolymeric polysaccharide, as a major virulence factor. However, the biosynthetic pathway of the EPS in the RSSC has not been fully characterized. To identify genes in EPS production beyond the EPS biosynthetic gene operon, we selected the EPS-defective mutants of R. pseudosolanacearum strain SL341 from Tn5-inserted mutant pool. Among several EPSdefective mutants, we identified a mutant, SL341P4, with a Tn5-insertion in a gene encoding a putative NDP-sugar epimerase, a putative membrane protein with sugar-modifying moiety, in a reverse orientation to EPS biosynthesis gene cluster. This protein showed similar to other NDP-sugar epimerases involved in EPS biosynthesis in many phytopathogens. Mutation of the NDP-sugar epimerase gene reduced EPS production and biofilm formation in R. pseudosolanacearum. Additionally, the SL341P4 mutant exhibited reduced disease severity and incidence of bacterial wilt in tomato plants compared to the wild-type SL341 without alteration of bacterial multiplication. These results indicate that the NDP-sugar epimerase gene is required for EPS production and bacterial virulence in R. pseudosolanacearum.

The Structure and Function of Locus Control Region (Locus Control Region의 구조와 기능)

  • Kim, Ae-Ri
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1587-1592
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    • 2007
  • Locus control region (LCR) is a cia-acting element which regulates the transcription of genes in developmental stage and/or tissue-specific pattern. Typically, LCR consists of several DNase I hypersensitive sites (HSs), where the binding motifs for transcriptional activators are present. The binding of activators to the HSs recruits chromatin modifying complexes to the LCR, opening chromatin structure and modifying histones covalently through the locus. LCR forms close physical contact with target gene located at a distance by looping away intervening region. In addition, non-coding RNA is transcribed from LCR toward target genes in continuously acetylated active domain. These structural and functional features of LCR suggest that the LCR plays many roles in chromatin activation and transcriptional regulation.

Production of miraculin protein in suspension cell lines of transgenic rice using Agrobacterium (Agrobacterium을 이용한 형질전환 벼 현탁 세포주에서 miraculin 단백질의 생산)

  • Kim, Hee Kyoung;Go, Ji Yun;Park, So-Young;Kang, Kwon Kyoo;Jung, Yu Jin
    • Journal of Plant Biotechnology
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    • v.47 no.3
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    • pp.227-234
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    • 2020
  • To produce the miraculin protein in suspension cultures, rice (Oryza sativa L.) was transformed with Agrobacterium tumefacience EHA105 containing the miraculin AB512278 gene. The cell suspension cultures were established using cell lines selected from transgenic rice callus. The integration of the miraculin gene into the rice chromosome was confirmed using genomic PCR analysis. In addition, RT-PCR analysis indicated that the miraculin gene is expressed in the selected suspension cell lines. Thus, the recombinant miraculin was expressed in the transgenic suspension cell line, HK-2. Therefore, we have successfully developed a HK-2 line that produces miraculin. These results demonstrate that transformed cell suspension cultures can be used to produce a taste-modifying protein such as miraculin.