• Title/Summary/Keyword: deactivated

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Determination of the Length of Target Recognition Sequence in sgRNA Required for CRISPR Interference (CRISPR 간섭에 필요한 sgRNA 표적 인식 서열 길이의 결정)

  • Kim, Bumjoon;Kim, Byeong Chan;Lee, Ho Joung;Lee, Sang Jun
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.534-542
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    • 2021
  • Single-molecular guide RNA (sgRNA) plays a role in recognizing the DNA target sequence in CRISPR technology for genome editing and gene expression control. In this study, we systematically compared the length of the target recognition sequence in sgRNAs required for genome editing using Cas9-NG (an engineered Cas9 recognizing 5'-NG as PAM sequence) and gene expression control using deactivated Cas9-NG (dCas9-NG) by targeting the gal promoter in E. coli. In the case of genome editing, the truncation of three nucleotides in the target recognition sequence (TRS) of sgRNA was allowed. In gene expression regulation, we observed that target recognition and binding were possible even if eleven nucleotides were deleted from twenty nucleotides of the TRS. When 4 or more nucleotides are truncated in the TRS of the sgRNA, it is thought that the sgRNA/Cas9-NG complex can specifically bind to the target DNA sequence, but lacks endonuclease activity to perform genome editing. Our study will be helpful in the development of artificial transcription factors and various CRISPR technologies in the field of synthetic biology.

Development of Reduced Graphene Oxide/Sr0.98Y0.08TiO3-δ Anode for Methane Fuels in Solid Oxide Fuel Cells (메탄연료사용을 위한 고체산화물 연료전지용 Reduced Graphene Oxide/Sr0.98Y0.08TiO3-δ 연료극 개발)

  • Hyung Soon Kim;Jun Ho Kim;Su In Mo;Gwang Seon Park;Jeong Woo Yun
    • Korean Chemical Engineering Research
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    • v.61 no.2
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    • pp.296-301
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    • 2023
  • Solid oxide fuel cell has received more attention recently due to the fuel flexibility via internal reforming. Commonly used Ni/YSZ anode, however, can be easily deactivated by carbon coking in hydrocarbon fuels. The carbon deposition problem can minimize by developing alternative perovskite anode. This study is focused on improving conductivity and catalytic activity of the perovskite anode by introducing rGO (reduced graphene oxide). Sr0.92Y0.08TiO3(SYT) anode with perovskite structure was synthesized with 1wt% of rGO. The presence of rGO during anode fabricating process and cell operation is confirmed through XPS and Raman analysis. The maximum power density of rGO/SYT anode improved to 3 times in H2 and 6 times in CH4 comparing to that of SYT anode due to the high electrical conductivity and good catalytic activity for CH4.

Similarities of Scaritoxin to Ciguatoxin on the Chromatographic Behaviours (Scaritoxin과 Ciguatoxin의 크로마토그라피상에서의 몇가지 유사성)

  • Joh, Yong-Goe;Scheuer, Paul J.
    • Korean Journal of Food Science and Technology
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    • v.17 no.2
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    • pp.121-127
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    • 1985
  • In studying the structural work on ciguatoxin, parrot fish collected were identified as Scarus sordidus, S. frenatus, S. scaber and S. pectarlis, in which only S. sordidus contained toxic materials. Crude toxins obtained by silicic acid column chromatography, could be separated on a DEAE-cellulose column into two fractions, ST-1(less polar) and ST-2(polar) eluted with chloroform and chloroform-methanol(1:1). Furthermore ST-1 could be changed into ST-2 by repeated chromatography on DEAE-cellulose. Rf values of ST-1 and ST-2 were 0.60-0.75 and 0.30-0.54 on TLC coated with silica gel 60F-254 developed by chloroform-methanol-water-acetic acid (90:9.5:0.2:0.3) mixture. The peaks of ST-1 and ST-2 were not observed on each HPLC chromatogram at low sensitivity(2X), but by bioassay they were detected in the fraction of 24-27ml(less polar toxin, 120ng) and 22-27 ml (polar toxin, 150 ng). Less polar ciguatoxin from morey eel viscera also showed its peak in the same elution volume(25ml). Being subjected to chromatography on basic aluminum oxide (activity grade I) or to alkaline treatment, followed by basic aluminum oxide (activity grade I) chromatography ST-1 toxin was remarkably converted into the polar toxic component supposed to be polar ciguatoxin in both cases. In the latter case, approximately 74% of the residual toxicity was changed into the polar component, accompanied by about 50% loss of the initial toxicity. More than 26% of ST-2 toxicity was transformed into the less polar toxic component supposed to be less polar ciguatoxin on a deactivated aluminum oxide (activity grade V) column.

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Scale-up Study of Heterogeneous Catalysts for Biodiesel Production from Nepalese Jatropha Oil (네팔산 자트로파 오일로부터 바이오디젤 제조를 위한 불균일계 촉매 Scale-up 연구)

  • Sim, Minseok;Lee, Seunghee;Kim, Youngbin;Ku, Huiji;Woo, Jaegyu;Joshi, Rajendra;Jeon, Jong-Ki
    • Clean Technology
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    • v.27 no.2
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    • pp.198-204
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    • 2021
  • This study focused on a two-step process using heterogeneous catalysts to produce biodiesel using Nepalese jatropha oil as a raw material. As a first step, the effect of the repetitive regeneration number of Amberlyst-15 on the esterification reaction of FFA in jatropha oil was investigated. Second, the possibility of a transesterification reaction scale-up using a dolomite bead catalyst was tested. Using 120 kg of jatropha seeds from Nepal, 30 L (27 kg) of jatropha oil was obtained, and the jatropha oil yield from the seeds was about 25.0 wt%. The acid value and FFA content of jatropha oil were measured to be 11.3 mgKOH g-1 and 5.65%, respectively. As a result of the esterification reaction of jatropha oil using the Amberlyst-15 catalyst in the form of beads, the acid value of the reaction product could be lowered to 0.26 mgKOH g-1 when the fresh Amberlyst-15 catalyst was used. As the regeneration of the Amberlyst-15 catalyst is repeated, the catalyst has been deactivated, and the esterification reaction performance has deteriorated. The cause of the deactivation seems to be due to the catalyst being broken and impurities being deposited. It was confirmed that the Amberlyst-15 catalyst could be reused up to 5 times for the esterification reaction of jatropha oil. In the second step, the transesterification reaction, a dolomite catalyst, was mass-produced and used in the form of beads. By transesterifying the pretreated jatropha oil in a spinning catalyst basket reactor equipped with 90 g of dolomite bead catalyst, 89.1 wt% of biodiesel yield was obtained in 2 hours after the start of the reaction, which was similar to the transesterification of soybean oil under the same conditions.