• 한국어병학회지
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• 제6권2호
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• pp.93-101
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• 1993

효모의 RNAI유전자는 RNA processing에 관여 하는지 혹은 RNA transport에 관여 하는지 아직까지 유전자의 기능이 정확히 알려져 있지 않은 실정이다. 효모의 RNAI 유전자의 기능을 파악하기 위한 방법으로 본 연구에서는 rna1-1 mutant gene을 cloning하여 이에 대한 DNA sequence를 조사함으로써 RNAI 유전자와 rna1-1 유전자의 차이점을 이해하고자 하였다. rna1-1 marker를 갖는 yeast strain(R49)로 부터 genomic DNA를 추출하여 이를 BglII로 절단하여 genomic southern blotting을 행한 결과 wild type의 경우와 동일하게 3.4 kb에서 hybridization되는 signal을 얻었으며, RNAl 및 rna1-1이 yeast genome내에 single site로 존재함을 보여 주는 결과를 얻었다. mutant strain으로 부터 얻은 3.4 kb의 BglI fragment를 pUC19의 BamHI site에 subcloning하여 transformant들을 얻었고, wild type RNAl 유전자를 probe로 하여 rna1-1 mutant 유전자를 cloning할 수 있었다. pUC19에 cloning된 RNA1유전자 및 rna1-1유전자로부터 다양한 Ba131유도체를 얻어 이들에 대한 염기 서열을 비교한 결과 transcription initation site에서부터 down stream쪽으로 17 아미노산위치에 TCC가 TTC로 대치되어 있었으며 그 결과 serine이 phenylalanine으로 변환되는 결과를 얻었다. Wild type RNAI gene의 5'-region에는 3군데의 TATA-like sequence가 true TATA box인지 확인하기 위하여 Bal3I deletion에 의해 -103nt까지 deletion된 유도체를 얻었으며 ${\Delta}RNAI$, rna1-1, 81-2-6 clone이 rna1-1 allele와 complementation한지 확인하였으나 ${\Delta}RNAI$은 TS-complementation을 하지 못하였다. 따라서 현재까지 TATA-box라고 알려진 부분은 promoter로 작용하지 못함을 확인하였다.

• 한국축산식품학회지
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• 제40권4호
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• pp.628-635
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• 2020

The objective of this study was to detect three non-halal meat products consisted of dog, pork, and rat species in meatball using novel multiplex-PCR with 12S rRNA gene as target sites. A total of 33 self-made meatballs were used, and they were grouped into eleven types of meatball based on meat species origin contained in the meatballs. Each type consisted of three meatballs. Extraction of genomic DNA from the meatballs was used as a DNA template for simplex-, duplex-, and multiplex-PCR processes. The result of simplex-PCR, duplex-PCR, and multiplex-PCR showed that the 12S rRNA primer gene successfully amplified DNA for each species bovine, dog, pig, and rat, which are respectively indicated by 155, 244, 357, and 491 bp of DNA bands. In addition, multiplex-PCR with 12S rRNA gene primers can be uniquely and accurately used for detection bovine, dog, pig, and rat species on beef meatball in one reaction.

• 대한미생물학회지
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• 제35권4호
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• pp.283-288
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• 2000

The PCR primer set JW21-JW22 of Weiss et al. (19), which was reported to amplify a 139-bp fragment of the l6S rRNA gene of Helicobacter pylori, has been recently used for the detection of H. pylori in clinical specimens. However, when we applied JW21-JW22 PCR to other members of the genus Helicobacter and unrelated microorganisms, all of these bacteria produced a 139-bp PCR product. Therefore, we designed a novel primer set, HPU185-HPL826, which produced a 642-bp amplicon of the l6S rRNA gene of H. pylori. Then we further examined the specificity of the novel PCR assay using Southern blot hybridization with an internal probe, HPP225. The PCR assay described in this study was shown to be highly sensitive and specific only to the H. pylori 16S rRNA gene sequences.

• 한국축산시설환경학회지
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• 제21권3호
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• pp.93-98
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• 2015

The objective of this study was to review application of next-generation sequencing (NGS) to investigate microbiome in the livestock sector. Since the 16S rRNA gene is used as a phylogenetic marker, unculturable members of microbiome in nature or managed environments have been investigated using the NGS technique based on 16S rRNA genes. However, few NGS studies have been conducted to investigate microbiome in the livestock sector. The 16S rRNA gene sequences obtained from NGS are classified to microbial taxa against the 16S rRNA gene reference database such as RDP, Greengenes and Silva databases. The sequences also are clustered into species-level OTUs at 97% sequence similarity. Microbiome similarity among treatment groups is visualized using principal coordinates analysis, while microbiome shared among treatment groups is visualized using a venn diagram. The use of the NGS technique will contribute to elucidating roles of microbiome in the livestock sector.

• 미생물학회지
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• 제27권2호
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• pp.77-84
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• 1989

Saccharomyces cerevisiae의 RNAI 유전자의 온도 감수성 돌연변이 균주는 성장허용 온도인 23"C에서는 정상적인 성"J--을하나, 성 장억제 온도인 36°C에서는 tRNA, rRNA 그러고 mRNA 의 선우울질을을 핵내에 축적함으후써 성장을 못한다. 본 실 험에서는 complementation 에 의하여 RNAI 유전자를 클로녕하였으며 concomitant loss 실험에 의하여 이 유천자의 클로닝 을 확인하였다. 유전자의 위치를 확인한 결과 3.5kb의 Bgl II 조각내에 RNAI 유전자가 존재함을 알 수 있였으며, 2.1kb에 해당하는 BamH I -Bgl II 조각에서는 RNAI 유전지에 의한 complementation 능력이 상실되는 것으후 보아 RNAI 유전자 는 3.5kb의 BglIl 조각내에 포함되는 BamH 1 자리 주위에 결쳐 있픔을 알 수 있었다.

• Mycobiology
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• 제33권2호
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• pp.109-112
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• 2005

The internal transcribed spacer (ITS) regions including the 3'-end of 18S rRNA gene, 5.8S rRNA gene and the 5'-end of the 28S rRNA gene of Rhizopus spp. were amplified by PCR and analyzed by DNASIS program. Length polymorphism of these region ranged from 564 bp in R. oryzae to 789bp in R. stolonifer. The length and sequence of 5.8S was very conserved with $154{\sim}155\;bp$. The sequence of ITS2 was more variable than that of ITS1. The base substitution rates were ranged from 0 to 0.6069 per site, and higher rate was found in R. stolonifer. In general, transition was usually more frequent than transversion. On the basis of sequencing results, four groups were clustered with value of 61.9% similarity; R. oryzae, R. micros pores, R. homothallicus, and R. stolonifer groups.

• 한국약용작물학회지
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• 제16권3호
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• pp.195-198
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• 2008

Karyotypic analysis and FISH (fluorescence in situ hybridization) with 45S and 5S rRNA genes were carried out in Persicaria tinctoria H Gross. The somatic metaphase chromosomes were ranged from 2.25 ${\mu}m$ to 1.50 ${\mu}m$ in length. Chromosome number was 2n = 4x = 40 with the basic number of x = 10. The chromosome complement of the species consisted of 16 pairs of metacentrics (chromososomes 1,2,3,4,6,7,8,9, 10, 11, 12, 13, 15, 18, 19 and 20) and 4 pairs of submetacentrics (chromosome 5, 14, 16 and 17). The karyotype formula was K(2n) = 4x = 32 m + 8 sm. In FISH analysis, three pairs of 45S rRNA gene loci on the terminal region of submetacentrics (chromosomes 5, 16 and 17) and two pairs of 5S rRNA gene loci on the centromeric region of metacentrics (chromosomes 9 and 11) were detected, respectively.

• Parasites, Hosts and Diseases
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• 제60권4호
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• pp.295-299
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• 2022

Malaria elimination and control require prompt and accurate diagnosis for treatment plan. Since microscopy and rapid diagnostic test (RDT) are not sensitive particularly for diagnosing low parasitemia, highly sensitive diagnostic tools are required for accurate treatment. Molecular diagnosis of malaria is commonly carried out by nested polymerase chain reaction (PCR) targeting 18S rRNA gene, while this technique involves long turnaround time and multiple steps leading to false positive results. To overcome these drawbacks, we compared highly sensitive cytochrome oxidase gene-based single-step multiplex reaction with 18S rRNA nested PCR. Cytochrome oxidase (cox) genes of P. falciparum (cox-III) and P. vivax (cox-I) were compared with 18S rRNA gene nested PCR and microscopy. Cox gene multiplex PCR was found to be highly specific and sensitive, enhancing the detection limit of mixed infections. Cox gene multiplex PCR showed a sensitivity of 100% and a specificity of 97%. This approach can be used as an alternative diagnostic method as it offers higher diagnostic performance and is amenable to high throughput scaling up for a larger sample size at low cost.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1288-1299
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• 2017

Composting is widely used to transform waste into valuable agricultural organic fertilizer. Anaerobic ammonium-oxidizing (anammox) bacteria play an important role in the global nitrogen cycle, but their role in composting remains poorly understood. In the present study, the community structure, diversity, and abundance of anammox bacteria were analyzed using cloning and sequencing methods by targeting the 16S rRNA gene and the hydrazine oxidase gene (hzo) in samples isolated from compost produced from cow manure and rice straw. A total of 25 operational taxonomic units were classified based on 16S rRNA gene clone libraries, and 14 operational taxonomic units were classified based on hzo gene clone libraries. The phylogenetic tree analysis of the 16S rRNA gene and deduced HZO protein sequences from the corresponding encoding genes indicated that the majority of the obtained clones were related to the known anammox bacteria Candidatus "Brocadia," Candidatus "Kuenenia," and Candidatus "Scalindua." The abundances of anammox bacteria were determined by quantitative PCR, and between $2.13{\times}10^5$ and $1.15{\times}10^6$ 16S rRNA gene copies per gram of compost were found. This study provides the first demonstration of the existence of anammox bacteria with limited diversity in cow manure composting.

• Archives of Pharmacal Research
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• 제27권2호
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• pp.225-231
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• 2004

This study was designed to investigate the effect of Trolox, a hydrophilic analogue of vitamin E, on the alteration of vasoregulatory gene expression during hepatic ischemia and reperfusion (I/R). Rats were subjected to 60 min of hepatic ischemia in vivo. The rats were treated intravenously with Trolox (2.5 mg/kg) or the vehicle as a control 5 min before reperfusion. Liver samples were obtained 5 h after reperfusion for a RT-PCR analysis on the mRNA for the genes of interest. These mRNA peptides are endothelin-1 (ET -1), potent vasoconstrictor peptide, its receptor $ET_A$ and $ET_B$, vasodilator endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and cyclooxygenase-2 (COX-2). It was seen that serum alanine aminotransferase and lipid peroxi-dation levels were markedly increased after I/R and Trolox significantly suppressed this increase. In contrast, the glutathione concentration decreased in the I/R group, and this decrease was inhibited by Trolox. ET-1 mRNA expression was increased by I/R, an increase which was prevented by Trolox. The mRNA levels for $ET_A$ receptor was significantly decreased, whereas ET$_{B}$ receptor transcript increased in the I/R group. The increase in $ET_A$ was prevented by Trolox. The mRNA levels for iNOS and HO-1 significantly increased in the I/R group and Trolox attenuated this increase. There were no significant differences in eNOS mRNA expression among any of the experimental groups. The mRNA levels for COX-2 and TNF-$\alpha$ significantly increased in I/R group and Trolox also attenuated this increase. Our findings suggest that I/R induces an imbalanced hepatic vasoregulatory gene expression and Trolox ameliorates this change through its free radical scavenging activity.y.