• Journal of Veterinary Science
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• v.23 no.2
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• pp.24.1-24.10
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• 2022

Background: Small interfering RNA technology has been considered a prospective alternative antiviral treatment using gene silencing against influenza viruses with high mutations rates. On the other hand, there are no reports on its effectiveness against the highly pathogenic avian influenza H5N1 virus isolated from Indonesia. Objectives: The main objective of this study was to improve the siRNA design based on the nucleoprotein gene (siRNA-NP) for the Indonesian H5N1 virus. Methods: The effectiveness of these siRNA-NPs (NP672, NP1433, and NP1469) was analyzed in vitro in Marbin-Darby canine kidney cells. Results: The siRNA-NP672 caused the largest decrease in viral production and gene expression at 24, 48, and 72 h post-infection compared to the other siRNA-NPs. Moreover, three serial passages of the H5N1 virus in the presence of siRNA-NP672 did not induce any mutations within the nucleoprotein gene. Conclusions: These findings suggest that siRNA-NP672 can provide better protection against the Indonesian strain of the H5N1 virus.

• 한국체육학회지인문사회과학편
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• v.55 no.4
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• pp.507-516
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• 2016

The purpose of this study is to identify the effects of PPARβ/δ over-expression on PGC-1α mRNA and protein stability after single bout of swimming exercise in mice skeletal muscle. Empty vector (EV) or PPARβ/δ was over-expressed in tibialis anterior(TA) using electroporation(EPO) technique to compare with non-treatment muscle(control; Con). TA muscles were dissected at 0h, 24h or 54h after termination of exercise. PGC-1α mRNA in Con, EV and PPARβ/δ over-expressed muscles were increased 6.8 fold (p<.001), 6.2 fold(p<.001) and 7.1 fold(p<.001), respectively, than sedentary(Sed) group at 0h after exercise and then reverted to Sed group levels at 24h and 54h after termination of exercise. PGC-1α and PGC-1α ubiquitination in EV treated muscles were increased 2.2 fold and 1.74 fold, respectively, than Sed group at 24h after termination of exercise, and then reverted to Sed group levels at 54h after termination of exercise. PGC-1α in PPARβ/δ over-expressed muscles at 24h and 54h after termination of exercise were increased 2.5 fold and 2.2 fold, respectively, than Sed group, but PGC-1α ubiquitination was not increased at 24h and 54h after termination of exercise. Our results indicate that PPARβ/δ over-expression does not increase PGC-1α mRNA stability, but increase PGC-1α protein stability through post-translation mechanism after termination of exercise.

• Microbiology and Biotechnology Letters
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• v.29 no.4
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• pp.234-239
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• 2001

In order to maximize the RNA accumulation and biomass production is Saccharomyces cerevisiae MTY62, a high-content RNA yeast strain, batch and fed-batch cultures were performed. Among the feeding modes of fed-batch cultures examined, the intermittent feeding mode R\`(IFB-lV), in which 50 ml of 40% molasses and 20% com steep liquor (CSL) solution was intermittently fed for 5 times, resulted in the cell concentration of 33.8 g- dry cell weight/1 and the RNA concentration of 5221 mg-/l, and RNA content of 153 mg-RNA/g-dry cell weight. The constant fed-batch with feeding mode III (CFB-III), in which the feeding rate of 40% molasses and 20% CSL solution was stepwisely decreased from 48 mph (9-13 h), to 24 mph (13-21 h), and to 18 ml/h (21∼ 48 h), gave the highest cell concentration of 42.7 g-dry ceil weigh71 and R7IA concentration of 5536 mg-RNA/1, which were about 2.4-fold and 1.9-fold increased levels, respectively, compared to the results of batch culture. However, the RNA con- tent of 130 mg-RNA/g-dry cell weight of the fed-batch was lower than that of the batch culture (171 mg-RNA/g-dry cell weight) and other fed-batch cultures. When the specific growth rates in the fed-batch cultures were increased, the RNA contents increased. This result indicates that the RNA content is adversely proportional to the cell concen- tration. However, at the same specific growth rate, the RNA content was maintained at higher level in the intermit- tent fed-batch than in the constant fed-batch culture.

• Journal of Food Hygiene and Safety
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• v.33 no.1
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• pp.71-76
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• 2018

Norovirus is a leading cause of sporadic pathogenic non-bacterial gastroenteritis worldwide. For the detection of norovirus, reverse transcription real-time PCR (RT qPCR) has quickly become a major tool due to its sensitivity and specificity. However, accurate viral RNA extraction methods are essential for RT qPCR analysis. TRIzol reagents are used to extract RNA from biological materials and are therefore widely used for norovirus RNA extraction. In this study, the yield of viral RNA extraction using TRIzol from genogroup II (GII) among the human norovirus genogroup I (GI) and GII, and murine norovirus (GV) depended on the pH of the virus sample solution. The yield of RNA extraction was higher at the alkaline pH than in the acidic region compared with the Ct (threshold cycle) value of the real-time PCR. From the results of this study, it was found that the pH condition is very important for the quantitative analysis of norovirus by extracting GII RNA using TRIzol.

• Korean Journal of Microbiology
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• v.26 no.1
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• pp.27-31
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• 1988

Biosynthesis and processing of cytoplasmic mRNA from heterogenous nuclear RNA (hn-RNA) in Aspergillus phoenicis were studied by $^{3}H$-uridine labeling and synchronous culture techniques during their life cycle. Incorporations of $^{3}H$-uridine into hn-RNA and mRNA were most rapid in vesicle-phialide fromation stage and diminished in hyphal growth stage. The processing of cytoplasmic mRNA from hn-RNA was proceeded more rapidly in hyphal growth and conidiophore formation stages than in conidia and vesicle-phialide formation stages. The specific radioactivities of hn-RNA and mRNA were very high in vesicle-phialide formation stage.

• Journal of Plant Biology
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• v.26 no.1
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• pp.1-6
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• 1983

In order to investigate the effects of GA3 on RNA biosynthesis, the amounts of rRNA and tRNA in germinating maize seeds were measured. The amount of rRNA in the endospermless seedlings was remarkably increased by GA3 tretment after 48 h of germination, but no effect was observed after 12 h of germination. While the amout of rRNA in 0.5 cm shoots in length was decreased by GA3 treatment, both of the amounts of rRNA and tRNA were increased in 1~1.5 cm shoots. According to the above mentioned results, it may be suggested that RNA biosynthesis is affected by GA3 treatment, and that GA3 participates in the biosynthesis of rRNA rather than tRNA in germinating maize seeds.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.68-72
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• 2002

To obtain a yeast mutant with high RNA content and high growth rate, Saccharomyces cerevisiae MTY62 was mutated with ethylmethane sulfonate. Among the selected mutants that were sensitive to the high concentration of KCl, M40-10 strain was finally selected due to its rapid cell growth and high RNA content in the tube and baffled-flask cultures. In the batch culture of M40-10 mutant, the maximum specific growth rate ($\mu_{max}$) of $0.38 h^{-1}$ , RNA concentration of 3210 mg-RNA/1, and RNA content of 183 mg-RNA/g-DCW were obtained, which were 23%, 15%, and 12% increased levels, respectively, compared to those of MTY62 parent strain. The intermittent fed-batch culture of M40-10 strain resulted in the maximum cell concentration of 35.6 g-DCW/1, RNA concentration of 5677 mg/1, and RNA content of 160 mg-RNA/g-DCW. Through the constant fed-batch culture, the maximum cell concentration of 46.4 g-DCW/1, RNA concentration of 6270 mg-RNA/1, and RNA content of 135 mg-RNA/g-DCW were obtained. At the 20 h culture time in the fed-batch cultures of M40-10 strain, the cell and RNA concentrations were increased by 30% and 10%, respectively, over the parent strain MTY62. In addition, it was also found that the accumulated RNA within the mutant cell was not degraded until the end of fed-batch cultivation, indicating that the M40-10 cell is a mutant with weak acidic RNase activity.y.

• Korean Journal of Food Science and Technology
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• v.27 no.1
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• pp.129-133
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• 1995

Continuous fermentations were performed in order to investigate the effect of culture condition on the yeast RNA accumulation and autolysis efficiency. The content of intracellular RNA increased with increasing dilution rate, showing its maximum value of 14.8% at D=0.35 $h^{-1}$. Also, both RNA productivity and specific RNA productivity tended to increase with the increase of dilution rate. The maximum biomass was obtained at $30^{\circ}C$ in the fixed dilution rate of 0.2 $h^{-1}$, whereas the maximum RNA content appeared at the lowest temperature experimented. Growth rate affected significantly on the yeast autolysis efficiency such that the extraction ratio(TN/TN) increased with increasing growth rate, whereas the hydrolysis ratio(AN/TN) was reversed. On the other hand, its efficiency was little affected by cultivation temperature.

• Korean Journal of Microbiology
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• v.27 no.2
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• pp.77-84
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• 1989

The temperature sensitive (ts) mutation on RNA1 gene of Saccharomyces cerevisiae prevents growth at restrictive temperature ($36^{\circ}C$) by accumulation of precursor tRNA, rRNA and mRNA (Hutchison et al., 1969; Shiokawa and Pogo, 1974; Hopper et al., 1978). RNA1 gene was cloned by complementation of the temperature sensitive growth defect of an rna1-1 mutant strain and identified by retransformation and concomitant loss of recombinant plasmid on non-selective condition. By deletion mapping, it was found that RNA1 gene resides within 3.5kb of BgII fragment.

• Journal of Microbiology and Biotechnology
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• v.19 no.8
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• pp.781-786
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• 2009

Small interfering synthetic double-stranded RNA (siRNA) was applied to suppress the expression of the human cytotoxic-T-Iymphocyte antigen 4-immunoglobulin (hCTLA4Ig) gene transformed in transgenic rice cell cultures. The sequence of the 21-nucleotide siRNA was deliberately designed and synthesized with overhangs to inactivate the expression of hCTLA4Ig. The chemically synthesized siRNA duplex was combined with polyethyleneimine (PEl) at a mass ratio of 1:10 (0.33 ${\mu}g$ siRNA:3.3 ${\mu}g$ PEl) to produce complexes. The siRNA complexes (siRNA+PEI) were labeled with Cy3 in order to subsequently confirm the delivery by fluorescent microscopy. In addition, the cells were treated with sonoporation at 40 kHz and 419W for 90 s to improve the delivery. The siRNA complexes alone inhibited the expression of hCTLA4Ig to 45% compared with control. The siRNA complexes delivered with sonoporation downregulated the production of hCTLA4Ig to 73%. Therefore, we concluded that the delivery of siRNA complexes into plant cells could be enhanced successfully by sonoporation.