• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.146-158
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• 2014

Camelina sativa that belongs to Brassicaceae family is an emerging oilseed crop. Camelina seeds contain approximately 40% storage oils per seed dry weight, which are useful for human and animal diets and industrial applications. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. In this study, three CsFAD2 genes (CsFAD2-1, CsFAD2-2 and CsFAD2-3.1) were isolated from developing seeds of Camelina sativa (L.) cv. CAME. The nucleotide and deduced amino acid sequences of three CsFAD2 genes were compared with those from dicotyledon and monocotyledon plants including Camelina cultivars Sunesone and SRS933. Three histidine motifs (HECGH, HRRHH, and HVAHH) required for FAD activity and a hydrophobic valine or isoleucine residue, which is a SNP (single nucleotide polymorphism) marker related with enzyme activity are well conserved in three CsFAD2s. The expressions of CsFAD2-1 and CsFAD2-3.1 were ubiquitously detected in various Camelina organs, whereas the CsFAD2-2 transcripts were predominantly detected in flowers and developing seeds. The contents of oleic acids decreased, whereas the amounts of linoleic acid increased in dry seeds of transgenic fad2-2 lines expressing each CsFAD2 gene compared with fad2-2 mutant, indicating that three CsFAD2 genes are functionally active. The isolated CsFAD2 genes might be applicable in metabolic engineering of storage oils with high oleic acids in oilseed crops.

• Plant Biotechnology Reports
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• v.4 no.3
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• pp.185-192
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• 2010

The Arabidopsis ${\omega}$-3 fatty acid desaturase (AtFAD7) catalyzes the synthesis of trienoic fatty acids (TA). A transgenic tobacco line, T15, was produced by a sense AtFAD7 construct and showed a cosuppression-like phenotype, namely extremely low TA levels. The sequence similarity between AtFAD7 and a tobacco ortholog gene, NtFAD7, was moderate (about 69%) in the coding sequences. AtFAD7 siRNAs accumulated at a high level, and both AtFAD7 and NtFAD7 mRNAs are degraded in T15 plants. The low-TA phenotype in T15 was dependent on a tobacco RNA-dependent RNA polymerase6 (NtRDR6). We also produced tobacco RNAi plants targeting AtFAD7 gene sequences. The AtFAD7 siRNA level was trace, which was associated with a slight reduction in leaf TA level. Unexpectedly, this RNAi plant showed an increased NtFAD7 transcript level. To investigate the effect of translational inhibition on stability of the NtFAD7 mRNAs, leaves of the wild-type tobacco plants were treated with a translational inhibitor, cycloheximide. The level of NtFAD7 mRNAs significantly increased after cycloheximde treatment. These results suggest that the translational inhibition by low levels of AtFAD7 siRNAs or by cycloheximide increased stability of NtFAD7 mRNA. The degree of silencing by an RNAi construct targeting the AtFAD7 gene was increased by co-existence of the AtFAD7 transgene, where NtRDR6-dependent amplification of siRNAs occurred. These results indicate that NtRDR6 can emphasize silencing effects in both cosuppression and RNAi.

• Journal of Veterinary Science
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• v.22 no.4
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• pp.42.1-42.16
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• 2021

Background: Inclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH. Objectives: To characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes. Methods: Suspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes. Results: The hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard. Conclusions: FAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.

• Korean Journal of Plant Tissue Culture
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• v.24 no.2
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• pp.93-97
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• 1997

For the molecular genetic study of cold tolerance mechanism in plants, a cDNA encoding fatty acid desaturase (fad3), converting linoleic acid (18:2, $\omega$-6) to linolenic acid (18:3, $\omega$-3), was isolated from $\lambda$ZAPII Arabidopsis thaliana cDNA expression library by plaque hybridization using fad3 cDNA probe derived from Brassica napus. A 1.8 kb-EcoRI fragment from a lambda clone showing a strong positive hybridization signal was subcloned into pGEM7 and analyzed for its nucleotide sequence. From deduced amino acid sequences, the fad3 gene was revealed to have an open reading frame(ORF) consisting of 386 amino acids with a molecular mass of 44,075 Da. The fad3 gene was compared to chloroplast $\omega$-3 fatty acid desaturase (fad7) and endoplasmic reticulum Δ12 fatty acid desaturase (fad2) to show 70% and 58% amino acid sequence homology, respectively, Especially, amino acids of internal (82 to 151) and carboxy terminal (276 to 333) regions were highly conserved, implying their requisite role for enzymatic functioning of fatty acid desaturases. IPTG-induced fad3 cDNA expression in E. coli cells was suggested to be toxic to bacterial growth.

• YAKHAK HOEJI
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• v.23 no.3_4
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• pp.147-152
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• 1979

Experiments were carried out to investigate for the interaction between FAD solution and synthetic resin containers made of polyvinylchloride(PVC), polyethylene(PE), and polycarbonate(PC), and for the effect of glycyrrhizine or malic acid on stabilization of FAD in aqueous solution by accelerated stability analysis. Analysis of FAD was determined by means of spectrometer and by separating by paper chromatography and metal ions were detected by atomic absorption spectrophotometer, which were extracted from containers by means of Food and Additive Regulation Standard. The thermal decomposition of FAD in aqueous solution was pseudo first order reaction and it was inhibited by adding glycyrrhizine or malic into the solution. PVC, PE and PC containers accelerated the decomposition of FAD in solution. It is assumed that bivalent heavy metals in resin containers may catalize the hydrolysis of FAD. The metals detected from the containers were Ca, Zn, Cu, Fe, Pb and Cd. And the total amounts of detected metals from PVC were 6.2mcg/cm$^{2}$, PE, 5.5mcg/cm$^{2}$, and PC, 2.7mcg/cm$^{2}$ which were proportional to the rate constant of FAD decomposition in aqueous solution.

• KSBB Journal
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• v.19 no.4
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• pp.331-334
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• 2004

In vivo characterization of FadB homologous enzymes including PaaG, YdbU and YgfG for medium-chain-length (MCL) polyhydroxyalkanoate (PHA) biosynthesis was carried out in fadB mutant Escherichia coli. Previously, it was reported that amplification of FadB homologous enzymes such as PaaG and YdbU in fadB mutant E. coli resulted in enhanced biosynthesis of MCL-PHA by greater than two fold compared with control strain. In this study, we constructed paaG fadB double mutant E. coli WB114 and ydbU fadB double mutant E. coli WB115 to investigate the roles of PaaG and YdbU in biosynthesis of MCL-PHA. Inactivation of paaG and ydbU genes in fadB mutant E. coli harboring Pseudomonas sp. 61-3 phaC2 gene reduced the MCL-PHA production to 0.16 and 0.16 PHA g/L, respectively from 2 g/L of sodium decanoate, which are much lower than 0.43 PHA g/L obtained with fadB mutant E. coli WB101 harboring the phaC2 gene. Also, we identified new FadB homologous enzyme YgfG, and examined its roles by overexpression of ygfG and construction of ygfG fadB double mutant E. coli WB113.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.281-286
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• 2005

We created a new graphite-Cu(II) electrode and found that the electrode could catalyze FADH$_2$ oxidation and FAD reduction coupled to electricity production and consumption, respectively. In a fuel cell with graphite-Cu(II) anode and graphite-Fe(III) cathode, the electricity was produced by coupling to the spontaneous oxidation of FADH$_2$ Fumarate and xylose were not produced from the enzymatic oxidation of succinate and xylitol without FAD, respectively, but produced with FAD. The production of fumarate and xylose in the reactor with FAD electrochemically regenerated was maximally 2- 5 times higher than that in the reactor with FAD. By using this new electrode with catalytic function, a bioelectrocatalysts can be engineered; namely, oxidoreductase (e.g., lactate dehydrogenase) and FAD can function for biotransformation without an electron mediator and second oxidoreductase for cofactors recycling.

• Journal of the Korean Society of Fisheries and Ocean Technology
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• v.56 no.4
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• pp.277-291
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• 2020

In order to understand characteristics on bycatch of Korean tuna purse seine fishery, especially operations associated with Fish Aggregating Device (FAD) in the Indian Ocean, we conducted analyses related to bycatch by school association type (unassociated school, FAD associated school and log associated school) using the data collected by scientific observers from 2016 to 2018. The FAD used by Korean tuna purse seine fishery in the Indian Ocean was a drifting FAD, which belongs to non-entangling FADs according to the category proposed by the International Seafood Sustainability Foundation (ISSF). The target species of Korean tuna purse seine fishery are skipjack, yellowfin and bigeye tunas, accounting for 99% of the total catch. The ratio of bycatch was 0.97% in total catch and the discard accounted for less than 1%, indicating that most catch was retained on board. In terms of bycatch ratio by school association type, it accounted for 0.12% for unassociated school, 1.09% for FAD associated school and 1.25% for log associated school. As for the catch proportion of shark species by school association type, it accounted for 0.01% for unassociated school, 0.11% for FAD associated school and 0.10% for log associated school, which showed that unassociated school type was the lowest to affect bycatch of non-target and shark species. Given the proportion of bycatch compositions, however, it is considered that FAD associated school of Korean tuna purse seine fishery has less caught bycatch species of non-target and shark, compared to other fleets operating in the Indian Ocean.

• Journal of Sensor Science and Technology
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• v.27 no.3
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• pp.156-159
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• 2018

We developed a glucose sensor using glucose dehydrogenase flavin adenine dinucleotide (GDH-FAD). The structure of the three-layer glucose sensor was simplified, in which a single-layer design was used to lower the unit cost, and GDH-FAD was used to increase the measurement reliability. GDH-FAD has less impact on the 20 interfering substances that affect blood glucose measurement, such as galactose and maltose compared to glucose oxidase (GOD), and is not affected by the oxygen saturation; therefore, it is possible to measure both arterial or venous blood and thus less susceptibility to hematocrit. In this study, we developed a single-layer glucose sensor strip with low hematocrit effect using the GDH-FAD enzyme, and measured and evaluated the performance.

• Proceedings of the KSME Conference
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• 2004.04a
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• pp.54-59
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• 2004

In this paper probabilistic fracture mechanics(PFM) approach is employed to evaluate the integrity of CANDU Zr-2.5Nb pressure tubes. Modified failure assessment diagram(Jr-FAD), plastic collapse, and critical crack length(CCL) approach are used for evaluating failure probability of the tubes. Jr-FAD was extended from the Kr-FAD because fracture of pressure tubes occurs in brittle manner due to hydrogen embrittlement of material by deuterium fluence. For developing the probabilistic integrity evaluation module, AECL procedures and fracture toughness parameters of EPRI were used.