• Title/Summary/Keyword: 6S RNA

Search Result 1,718, Processing Time 0.03 seconds

RNA Production from the Hydrolyzate of Sliced and Dried Sweet Potatoes. (절간고구마를 이용한 RNA 생산에 관한 연구)

  • 이종임;송재철;조원대;양한철
    • Microbiology and Biotechnology Letters
    • /
    • v.8 no.1
    • /
    • pp.9-18
    • /
    • 1980
  • During an extensive screening tests of yeasts for their RNA formation, it was found that Cryptococcus laurentii had especially high RNA content and high dry cell weight, when hydrolyzate of sliced and dried sweet potatoes was used as a carbon source. Growth conditions of this strain were examined, and the most desirable results were obtained at 48 hours of cultivation on a reciprocal shaker at 3$0^{\circ}C$ with initial pH 6.0. Under the above conditions, the RNA content and yield of dry cells were investigated using various media compositions. Ammonium sulfate 0.40%, peptone 0.6 %, and yeast extract 0.4% were appeared to be favorable as a nitrogen sources. The optimum concentrations of K $H_2$P $O_4$, M $n^{++}$, C $O^{++}$ were 0.05 %, 0.1 %, and 0.001 %, respectively. Ca-pantothenate, 400$\mu\textrm{g}$/$m\ell$, showed relatively favorable effects as a growth factor. The maximum RNA content obtained in this study was 16.8 % of the total dry cell weight.t.t.

  • PDF

Improving amber suppression activity of an orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase and a variant of E. coli initiator tRNA, fMam tRNACUA, for the efficient incorporation of unnatural amino acids (효율적인 비천연 아민노산 도입을 위한 효모균 타이로신-tRNA 합성효소와 대장균 시작 tRNA 변이체의 엠버써프레션 활성증가)

  • Tekalign, Eyob;Oh, Ju-Eon;Park, Jungchan
    • Korean Journal of Microbiology
    • /
    • v.54 no.4
    • /
    • pp.420-427
    • /
    • 2018
  • The orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase (Sc YRS) and a variant of E. coli initiator tRNA, fMam $tRNA_{CUA}$ which recognizes the amber stop codon is an effective tool for site-specific incorporation of unnatural amino acids into the protein in E. coli. To evolve the amber suppression activity of the orthogonal pair, we generated a mutant library of Sc YRS by randomizing two amino acids at 320 and 321 which involve recognition of the first base of anticodon in fMam $tRNA_{CUA}$. Two positive clones are selected from the library screening with chloramphenicol resistance mediated by amber suppression. They showed growth resistance against high concentration of chloramphenicol and their $IC_{50}$ values were approximately 1.7~2.3 fold higher than the wild type YRS. In vivo amber suppression assay reveals that mutant YRS-3 (mYRS-3) clone containing amino acid substitutions of P320A and D321A showed 6.5-fold higher activity of amber suppression compared with the wild type. In addition, in vitro aminoacylation kinetics of mYRS-3 also showed approximately 7-fold higher activity than the wild type, and the enhancement was mainly due to the increase of tRNA binding affinity. These results demonstrate that optimization of anticodon recognition by engineered aminoacyl tRNA synthetase improves the efficiency of unnatural amino acid incorporation in response to nonsense codon.

The Effects of Hibiscus syriacus(HSS) Extract on the Alzheimer's Disease Model Induced by CT-105 and ${\beta}A$ (목근피(木槿皮)가 CT105와 ${\beta}A$로 유도된 Alzheimer's Disease 병태(病態) 모델에 미치는 영향)

  • Choi, Byung-Man;Jung, In-Chul;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
    • /
    • v.15 no.2
    • /
    • pp.119-139
    • /
    • 2004
  • This research investigates the effect of the Hibiscus syriacus(HSS) on Alzheimer's disease. Specifically, the effects of the HSS extract on (1) $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ mRNA of PC-12 cells treated with LPS; (2) amyloid precursor proteins(APP), acetylcholinesterase(AChE), and glial fibrillary acidic protein(GFAP) mRNA of PC-12 cells treated with CT-105; (3) the AChE activity and the APP production of PC-12 cell treated with CT-105; (4) the behavior; (4) expression of $IL-1{\beta}$, $TNF-{\alpha}$, $IL-1{\beta}$ mRNA, $TNF-{\alpha}$ mRNA, and reactive oxygen species(ROS); (5) the infarction area of the hippocampus, and brain tissue injury in Alzheimer's diseased mice induced with ${\beta}A$ were investigated. The results were summarized below ; 1. The HSS extract suppressed the expression of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ mRNA in THP-l cells treated with LPS. 2. The HSS extract suppressed the expression of APP, AChE, and GFAP mRNA in PC-12 cells treated with CT-105. 3. The HSS extract suppressed the AChE activity, and the production of APP significantly in PC-12 cells treated with CT-105. 4. For the HSS extract group a significant inhibitory effect on the memory deficit was shown for the mice with Alzheimer's disease induced by ${\beta}A$ in the Morris water maze experiment, which measured stop-through latency, and distance movement-through latency. 5. The HSS extract suppressed the over-expression of $IL-1{\beta}$, $TNF-{\alpha}$, $IL-1{\beta}$ and $TNF-{\alpha}$ mRNA, CD68/GFAP, ROS in the mice with Alzheimer's disease induced by ${\beta}A$. 6. The HSS extract reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by ${\beta}A$. These results suggest that the HSS extract may be effective for the prevention and treatment of Alzheimer's disease. Investigation into the clinical use of the HSS extract for Alzheimer's disease is suggested for future research.

  • PDF

Isolation and Characterization of Pre-$tRNA^{Val}$ Splicing Mutants of Schizosaccharomyces pombe

  • Hwang, Ku-Chan;Kim, Dae-Myung
    • Journal of Microbiology
    • /
    • v.35 no.4
    • /
    • pp.334-340
    • /
    • 1997
  • A collection of 132 temperature sensitive (ts) mutants was generated by the chemical mutagenesis of Schizosaccharomyces pombe wild type strain and screened for tRNA splicing defects on Northern blots by hybridization with an oligonucleotide that recognizes the exon of the S. pombe tRNA^Val as a probe. We identidied 6 mutants which accumulate precursor $tRNA^{Val}$. Among them, 2 mutants exhibited remarkable morphological differences compared to wild type cells. One tRNA splicing mutant showed elongated cell shape in permissive as well as non-permissive cultures. The other mutant exhibited shortened cell morphology only in nonpermissive culture. The total RNA pattern in the splicing mutants appeared to be normal. Genetic analysis of four $tRNA^{Val}$ splicing mutants demonstrated that the mutation reside in different genes.

  • PDF

Construction of recombinant DNA clone for bovine viral diarrhea virus (소 바이러스성 설사병 바이러스의 유전자 재조합 DNA clone의 작성에 관한 연구)

  • Yeo, Sang-geon;Cho, H.J.;Masri, S.A.
    • Korean Journal of Veterinary Research
    • /
    • v.32 no.3
    • /
    • pp.389-398
    • /
    • 1992
  • Molecular cloning was carried out on the Danish strain of bovine viral diarrhea virus(BVDV) to construct strategy for the diagnostic tools and effective vaccine of BVD afterwards. A recombinant DNA clone(No. 29) was established successfully from cDNA for viral RNA tailed with adenine homopolymer at 3'-end. $^{32}P$-labeled DNA probes of 300~1,800bp fragments, originating from the clone 29, directed specific DNA-RNA hybridization results with BVDV RNA. Recombinant DNA of the clone 29 was about 5,200bp representing 41.6% of the full length of Danish strain's RNA, and restriction sites were recognized for EcoR I, Sst I, Hin d III and Pst I restriction enzymes in the DNA fragment.

  • PDF

MicroRNA Profile in the Helicobacter pylori-infected Gastric Epithelial Cells (Helicobacter pylori 감염 위상피세포에서 MicroRNA 발현 변화)

  • Chang Whan Kim;Sung Soo Kim;Tae Ho Kim;Woo Chul Chung;Jae Kwang Kim
    • Journal of Digestive Cancer Research
    • /
    • v.5 no.2
    • /
    • pp.105-112
    • /
    • 2017
  • Background: The expression of miRNAs in response to Helicobacter pylori infection has not been well explored. The aims of this study were to evaluate the H. pylori associated miRNAs in the gastric epithelial cells. Methods: We investigated gastric epithelial cell-line (HS3C) exposed H. pylori over 3 months and AGS cell-line (AGS) exposed H. pylori for 6 hour. After the extraction of miRNA from these cell-lines, microarray and real time PCR were performed to confirm the alteration of expression. Results: All 12 miRNAs chosen for real-time PCR are based on the result of microarray and their potential functions related to H. pylori infection. miR-21, miR-221, miR-222 were upregulated in the H. pylori infected AGS cell for 6 hours and HS3C cells. miR-99b, miR-200b, miR-203b and miR-373 were downregulated in the H. pylori infected AGS cell for 6 hours and HS3C cells. miR-23a, miR-23b, miR-125b, miR-141 and miR-155 were upregulated in HS3C cell line but not in H. pylori infected AGS cell for 6 hours. Conclusion: miR-21, miR-99b, miR-125b, miR-200b, miR-203b, miR-221, miR-222, and miR-373 are supposed to be related with oncogenesis of H. pylori infection. Further studies are needed for the evaluation of the function of these confirmed miRNAs.

  • PDF

Acinetobacter marinus sp. novo and Acinetobacter seohaensis sp. nov., Isolated from Sea Water of the Yellow Sea in Korea

  • Yoon, Jung-Hoon;Kim, In-Gi;Oh, Tae-Kwang
    • Journal of Microbiology and Biotechnology
    • /
    • v.17 no.11
    • /
    • pp.1743-1750
    • /
    • 2007
  • Two Gram-negative, nonmotile, coccobacilli, SW-$3^T$ and SW-$100^T$, were isolated from sea water of the Yellow Sea in Korea. Strains SW-$3^T$ and SW-$100^T$ contained ubiquinone-9 (Q-9) as the predominant respiratory lipoquinone and $C_{18:1}\;{\omega}9c$ and $C_{16:0}$ as the major fatty acids. The DNA G+C contents of strains SW-$3^T$ and SW- $100^T$ were 44.1 mol% and 41.9 mol%, respectively. A neighbor-joining tree based on l6S rRNA gene sequences showed that the two isolates fell within the evolutionary radiation enclosed by the genus Acinetobacter. Strains SW-$3^T$ and SW-$100^T$ exhibited a l6S rRNA gene similarity value of 95.7% and a mean DNA-DNA relatedness level of 9.2%. Strain SW-$3^T$ exhibited l6S rRNA gene sequence similarity levels of 93.5-96.9% to the validly described Acinetobacter species and fifteen Acinetobacter genomic species. Strain SW-$100^T$ exhibited l6S rRNA gene sequence similarity levels of less than 97.0% to the other Acinetobacter species except Acinetobacter towneri DSM $14962^T$ (98.0% similarity). Strains SW-$3^T$ and SW-$100^T$ exhibited mean levels of DNA-DNA relatedness of 7.3-l6.7% to the type strains of some phylogenetically related Acinetobacter species. On the basis of phenotypic, phylogenetic, and genetic data, strains SW-$3^T$ and SW-$100^T$ were classified in the genus Acinetobacter as two distinct novel species, for which the names Acinetobacter marinus sp. novo (type strain SW-$3^T$=KCTC $12259^T$=DSM $16312^T$) and Acinetobacter seohaensis sp. novo (type strain SW-$100^T$=KCTC $12260^T$=DSM $16313^T$) are proposed, respectively.

Taxonomic characteristics of novel Flavobacteriumsp. B1 from a freshwater pond

  • Bae, Young-Min
    • Journal of the Korean Applied Science and Technology
    • /
    • v.39 no.5
    • /
    • pp.605-613
    • /
    • 2022
  • The genus Flavobacterium, type genus of the family Flavobacteriaceae and a member of the phylum Bacteriodetes includes gram-negative and yellow-pigmented rods. Those bacteria have been isolated from various environments of the earth. A yellow-pigmented, gram-negative rod was isolated from a pond in the campus of the Changwon University, Changwon, Kyeongnam and designated as strain B1. Strain B1 was further analyzed physiologically, biochemically and phylogenetically, and concluded to be a member of genus Flavobacterium. BLAST search of the 16S rRNA gene sequence of strain B1 shows homology no higher than 99.0% with those sequences of other bacteria. The major fatty acids of strain B1 are iso-C15:0 (19.6%), summed feature 3(C16:1 ω7c and/or C16:1 ω6c, 16.1%), iso-C17:0 3OH(10.2%), iso-C15:0 3OH(8.4%) and iso-C15:1 G(6.6%) showing significant differences in fatty acid compositions between strain B1 and the other known Flavobacterium species. DNA sequence of 16S rRNA gene of strain B1 was deposited in genbank under accession number OP060681.

Taxonomic characterization of novel Hymenobacter sp. B2 isolated from a freshwater environment (민물환경에서 분리된 novel Hymenobacter sp. B2의 분류학적 특성연구)

  • Young-Min Bae
    • Journal of the Korean Applied Science and Technology
    • /
    • v.40 no.4
    • /
    • pp.881-889
    • /
    • 2023
  • The genus Hymenobacter, type genus of the family Hymenobacteraceae and a member of the phylum Bacteroidota includes gram-negative and red-pigmented rods. Those bacteria have been isolated from various environments of the earth. I isolated a red-pigmented, gram-negative rod from a pond in the campus of the Changwon University, Changwon, Kyeongnam and designated this bacterium as strain B2. Strain B2 was further analyzed phylogenetically and biochemically, and concluded as a member of genus Hymenobacter. BLAST search of the 16S rRNA gene sequence of strain B2 showed its homology lower than 98.7% with those sequences of the other bacteria whose 16S rRNA gene sequences have been reported. Fatty acid composition of the strain B2 was analyzed and its major fatty acids are summed feature 3(C16:1 ω7c and/or C16:1 ω6c, 22.8%), iso-C15:0 (16.2%), anteiso-C15:0(12.9%), C16:1ω5c(12.4%) and summed feature 4 (iso-C17:1 I/anteiso-C17:1)(9.5%) showing significant differences in fatty acid compositions between strain B2 and the other known Hymenobacter species. DNA sequence of 16S rRNA gene of strain B2 was deposited in genbank under accession number OQ318247.

Comparison of Culture-dependent and DGGE based Method for the Analysis of Marine Bacterial Community (배양법과 DGGE에 의한 해양세균 군집의 비교분석)

  • Kim, Mal-Nam;Bang, Hyo-Joo
    • Korean Journal of Environmental Biology
    • /
    • v.24 no.4
    • /
    • pp.307-313
    • /
    • 2006
  • Seasonal variation of marine bacterial community was analyzed in the surface sea water collected from one of the stations locating at Tongyeoung coastal area, Korea. The results obtained by the culture method through identification with the VITEK Microbe ID system after pure culture in the selective medium were compared with those obtained by the DGGE based 16S rRNA PCR method. The composition of the marine bacterial community in the sea water samples harvested in September, 2004, November, 2004, January, 2005, May, 2005 and August, 2005 determined by the culture method showed 5, 5, 4, 6, and 10 strains respectively. Pseudomonas fluorescens and Acinetobacter lwoffii were detected in all seasons. The other strains were identified to be Pseudomonas stutzeri, Sphingomonas paucimobilis, Burkholderia mallei and Chryseobacterium indologenes. In contrast, the 16S rRNA PCR-DGGE method detected 10, 11, 6, 9 and 13 populations respectively in the same sea water samples and the strains were identified to be Acinetobacter lwoffii, Burkholderia mallei, Pseudomonas fluoresence, Actinobacillus ureae, Burkholderia sp., Pseudomonas stutzeri, Roseobacter sp., Vibrio parahaemolyticue, Sphingomonas paucimobilis and Rugeria algocolus. This results indicated that the DGGE based 16S rRNA PCR method was more efficient than the culture method for the grasp of the characteristics of the marine bacterial community.