• Title/Summary/Keyword: antisense

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Molecular Characterization of an Isolate of Bean Common Mosaic Virus First Identified in Gardenia Using Metatranscriptome and Small RNA Sequencing

  • Zhong-Tian Xu;Hai-Tao Weng;Jian-Ping Chen;Chuan-Xi Zhang;Jun-Min Li;Yi-Yuan Li
    • The Plant Pathology Journal
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    • v.40 no.1
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    • pp.73-82
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    • 2024
  • Gardenia (Gardenia jasminoides) is a popular and economically vital plant known for its ornamental and medicinal properties. Despite its widespread cultivation, there has been no documentation of plant viruses on gardenia yet. In the present study, gardenia leaves exhibiting symptoms of plant viral diseases were sampled and sequenced by both metatranscriptome and small RNA sequencing. As a consequence, bean common mosaic virus (BCMV) was identified in gardenia for the first time and named BCMV-gardenia. The full genome sequence of BCMV-gardenia is 10,054 nucleotides (nt) in length (excluding the poly (A) at the 3' termini), encoding a large polyprotein of 3,222 amino acids. Sequence analysis showed that the N-termini of the polyprotein encoded by BCMV-gardenia is less conserved when compared to other BCMV isolates, whereas the C-termini is the most conserved. Maximum likelihood phylogenetic analysis showed that BCMVgardenia was clustered closely with other BCMV isolates identified outside the leguminous plants. Our results indicated that the majority of BCMV-gardenia virus-derived small interfering RNAs (vsiRNAs) were 21 nt and 22 nt, with 21 nt being more abundant. The first nucleotide at the 5' termini of vsiRNAs derived from BCMV-gardenia preferred U and A. The ratio of vsiRNAs derived from sense (51.1%) and antisense (48.9%) strands is approaching, and the distribution of vsiRNAs along the viral genome is generally even, with some hot spots forming in local regions. Our findings could provide new insights into the diversity, evolution, and host expansion of BCMV and contribute to the prevention and treatment of this virus.

MOLECULAR BIOLOGIC ANALYSIS OF FMR-1 GENE TRINUCLEOTIDE REPEATS IN AUTISTIC PATIENTS (자폐장애 환자에서 FMR-1 유전 삼염기 반복의 분자생물학적 분석)

  • Kwak, Ho-Soon;Chun, Hyo-Jin;Chang, Eun-Jin;Kim, Hee-Cheol;Kim, Jung-Bun;Park, Young-Nam;Jung, Chul-Ho
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.11 no.1
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    • pp.3-15
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    • 2000
  • Objectives:There has been a rapid expansion of studies aimed at elucidating the genetic basis of autistic disorder, especially it’ relationship to fragile-X syndrome. The detection of fragile X chromosome(Xq27.3) by cytogenetic analysis has revealed many difficulties in testing. Therefore, to explore the relationship between autistic disorder and fragile X syndrome, this study administered molecular biologic methods which examined an unstable CGG repeat within the fragile X mental retardation-1(FMR-1) gene. Methods:Ninety nine autistic children and eight normal control children were tested. The number of CGG repeats within FMR-1 gene was measured after amplification by PCR, and cytogenetic analysis was also carried out to detect fragile site Xq27.3. Southern blot hybridization, using StB12.3 and/or Pfxa3 probe, was done for the patients showing expansion of more than 50 CGG repeats (premutation). Results:All but two autistic patients had no expansion in CGG repeats by PCR and there was no significant statistical difference in number of CGG repeat in comparison with normal control. Two autistic patients, considered as premutation by PCR analysis, had no full mutation or premutation by Southern blot hybridization. All autistic children tested did not have any abnormal karyotype or fragile site Xq27.3. Conclusions:These results suggest that autistic patients may not have abnormality in FMR-1 gene or abnormal expansion in CGG repeat. In conclusion, fragile X syndrome may not be antecedent of autistic disorder.

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Characterization of Chemical Composition in Poplar wood (Populus deltoides) by Suppression of CCoAOMT Gene Expression (CCoAOMT 유전자 발현 억제에 의한 현사시나무의 화학조성 변화)

  • Eom, In-Yong;Kim, Kwang-Ho;Lee, Soo-Min;Yi, Yong-Sub;Choi, Joon-Weon
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.3
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    • pp.213-222
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    • 2010
  • In this study, chemical compositions - holocellulose, lignin and monomeric sugars - were characterized with two poplar wood cell walls, one of which was grown at normal condition (CPW) and the other was genetically modified by antisence suppression of CCoAOMT gene expression (ACPW). Milled wood lignins were isolated from CPW and ACPW and subjected to methoxyl group, DFRC, Py-GC/MS, GPC, $^{13}C$-NMR analysis, respectively. There were few differences in holocellulose contents in both cell walls, which were determined to 81.6% in CPW and to 82.3% in ACPW. However, lignin contents in ACPW was clearly decreased by the suppression of CCoAOMT gene expression. In CPW 21.7% of lignin contents was determined, while lignin contents in ACPW was lowered to 18.3%. The relative poor solubility of ACPW in alkali solution could be attributed to the reduction of lignin content. The glucose contents of CPW and ACPW were measured to 511.0 mg/g and 584.8 mg/g and xylose contents 217.8 mg/g and 187.5 mg/g, respectively, indicating that suppression of CCoAOMT gene expression could be also influenced to the formation of monomeric sugar compositions. In depth investigation for milled wood lignin (MWL) isolated from both samples revealed that the methoxyl contents at ACPW was decreased by 7% in comparison to that of CPW, which were indirectly evidenced by $^{13}C$-NMR spectra and Py-GC/MS. According to the data from Py-GC/MS S/G ratios of lignin in CPW and ACPW were determined to 0.59 and 0.44, respectively. As conclusive remark, the biosynthesis of syringyl unit could be further influenced by antisense suppression of CCoAOMT during phenylpropanoid pathway in the plant cell wall rather than that of guaiacyl unit.

Flavonoid Biosynthesis: Biochemistry and Metabolic Engineering (Flavonoid 생합성:생화학과 대사공학적 응용)

  • Park, Jong-Sug;Kim, Jong-Bum;Kim, Kyung-Hwan;Ha, Sun-Hwa;Han, Bum-Soo;Kim, Yong-Hwan
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.265-275
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    • 2002
  • Flavonoid biosynthesis is one of the most extensively studied areas in the secondary metabolism. Due to the study of flavonoid metabolism in diverse plant system, the pathways become the best characterized secondary metabolites and can be excellent targets for metabolic engineering. These flavonoid-derived secondary metabolites have been considerably divergent functional roles: floral pigment, anticancer, antiviral, antitoxin, and hepatoprotective. Three species have been significant for elucidating the flavonoid metabolism and isolating the genes controlling the flavonoid genes: maize (Zea mays), snapdragon (Antirrhinum majus) and petunia (Prtunia hybrida). Recently, many genes involved in biosynthesis of flavonoid have been isolated and characterized using mutation and recombinant DNA technologies including transposon tagging and T-DNA tagging which are novel approaches for the discovery of uncharacterized genes. Metabolic engineering of flavonoid biosynthesis was approached by sense or antisense manipulation of the genes related with flavonoid pathway, or by modified expression of regulatory genes. So, the use of a variety of experimental tools and metabolic engineering facilitated the characterization of the flavonoid metabolism. Here we review recent progresses in flavonoid metabolism: confirmation of genes, metabolic engineering, and applications in the industrial use.

Relationships of the Lithium-Induced Growth Inhibition of C6 Rat Glioma Cell to Expression of the Insulin-like Growth Factor System Components (C6 Rat Glioma Cell에서 리튬에 의한 성장 억제와 Insulin-like Growth Factor System Components의 발현과의 관계)

  • Kim, I.A.;Jin, E.J.;Cho, E.J.;Sohn, S.H.;Lee, C.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.563-570
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    • 2004
  • The insulin-like growth factor(IGF) system, consisting of IGFs-I and -II ligands and their receptors and six IGF-binding proteins(IGFBPs), plays an important role in survival, proliferation and differentiation of a variety of cell types. Lithium is a known modulator of survival and proliferation of many cell types in vitro. The present study was undertaken to investigate the relationship between LiCI-induced changes in cell survival and growth and the expression of the IGF system components in C6 rat glioma cell line which, besides IGF-I and its receptor, is known to express IGFBP-3 as its major IGF carrier. When C6 cells were cultured for 24h in the absence or presence of 2mM or 5mM LiCl in a 10% serwn-containing medium, the viability and the number of cells were not affected by added lithium. In 72-h culture, however, C6 cells clearly exhibited a dose-dependent response to added LiCl. The cells cultured for 72h in the presence of 0, 2mM and 5mM LiCl exhibited a typical mitotic, a growth-arrested and an apoptotic appearances, respectively. Moreover, the apoptotic cells were accompanied by reduced expression of IGF-I, IGF-I receptor and IGFBP-3 as examined by semi-quantitative reverse transcription-polymerase chain reaction. Interestingly, blockade of IGFBP-3 mRNA translation by addition of 101${\mu}M$ IGFBP-3 anti-sense oligodeoxyribonucleotide in serum-free, 24-h culture resulted in a decrease in the number of cells as well as relative abundance of the target mRNA. In summary, results suggest that the cytotoxic effect of lithium in C6 cell is likely to be mediated, in part, by suppression by this agent of the expression of the IGF system components. In this regard, IGFBP-3 may play at least a 'permissive' role in normal proliferation of this cell.