• Genomics & Informatics
• /
• v.12 no.2
• /
• pp.71-75
• /
• 2014

The tRNA structure contains conserved modifications that are responsible for its stability and are involved in the initiation and accuracy of the translation process. tRNA modification enzymes are prevalent in bacteria, archaea, and eukaryotes. tRNA Gm18 methyltransferase (TrmH) and tRNA $m^1G37$ methyltransferase (TrmD) are prevalent and essential enzymes in bacterial populations. TrmH involves itself in methylation process at the 2'-OH group of ribose at the 18th position of guanosine (G) in tRNAs. TrmD methylates the G residue next to the anticodon in selected tRNA subsets. Initially, $m^1G37$ modification was reported to take place on three conserved tRNA subsets ($tRNA^{Arg}$, $tRNA^{Leu}$, $tRNA^{Pro}$); later on, few archaea and eukaryotes organisms revealed that other tRNAs also have the $m^1G37$ modification. The present study reveals Gm18, $m^1G37$ modification, and positions of $m^1G$ that take place next to the anticodon in tRNA sequences. We selected extremophile organisms and attempted to retrieve the $m^1G$ and Gm18 modification bases in tRNA sequences. Results showed that the Gm18 modification G residue occurs in all tRNA subsets except three tRNAs ($tRNA^{Met}$, $tRNA^{Pro}$, $tRNA^{Val}$). Whereas the $m^1G37$ modification base G is formed only on $tRNA^{Arg}$, $tRNA^{Leu}$, $tRNA^{Pro}$, and $tRNA^{His}$, the rest of the tRNAs contain adenine (A) next to the anticodon. Thus, we hypothesize that Gm18 modification and $m^1G$ modification occur irrespective of a G residue in tRNAs.

• Journal of radiological science and technology
• /
• v.21 no.1
• /
• pp.65-68
• /
• 1998

We attempted to purify radiolabeled RNA using Sephadex G-15 and G-50 chromatography instead of commercial RNA purification kit. In the Sephadex G-15 chromatography the major portion of RNA was eluted in the fractions ranging from 3rd to 5th whereas broad elution profile of RNA was obtained from the Sephadex G-50 chromatography. The elution profile and purity of RNA obtained from Sephadex G-15 chromatography was very similar to that by commercial RNA purification kit. Furthermore, operating time required for purification of RNA by Sephadex G-15 was rather smaller than that by commercial kit. Overall results suggest that the purification of radiolabeled RNA using Sephadex G-15 is more money and time saying than using commercial RNA purification kit.

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

• Journal of the Korean Chemical Society
• /
• v.46 no.2
• /
• pp.157-163
• /
• 2002

To identify RNA motifs interacting with $tRNA^{Val}$, a SELEX(Systematic Evolution of Ligands by Exponential Enrichment) was applied. Random DNA library which contains a region of ran-domized 48-mer oligonucleotide flanked by conserved sequ ence primers was transcribed into RNA pool using T7 RNA polymerase and RNA aptamers were selected with $tRNA^{Val}$ -immobilized affinity column through 14 rounds of SELEX. Some of the resulting aptamers contained a consensus sequence similar to the sequence in the loop regions of three rRNAs; C43GAAC47 sequence of 5S rRNA, G1491AAGU1495, G1379UUCC1383 sequence of 16S rRNA and C1064UUAG1068, G2110UGUA2114, C2480GACGG2485, A2600CAGU2604 sequence of 23S rRNA. These results suggest that $tRNA^{Val}$ can interact with 5S rRNA, 16S rRNA and 23S rRNA with variety in ribosome.

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

• Korean Journal of Microbiology
• /
• v.46 no.3
• /
• pp.229-232
• /
• 2010

Enolase is one of the glycolytic enzymes, which are involved in a central energy metabolism present in nearly all organisms. In Escherichia coli, enolase constitutes RNA degradosome with RNase E, PNPase and RNA helicase, which are involved in most mRNA degradation and RNA processing. Recently, it has been reported that RNase G, an RNase E homolog, degrades eno mRNA. To examine a functional role of RNase G in enolase expression which is known to be up-regulated under microaerobic condition, we carried out experiments. Here, we report that expression levels of enolase and RNase G are not correlated under microaerobic condition. Based on this observation, we suggest the existence of an unknown factor(s) which regulate the activity of RNase G or enolase mRNA under microaerobic conditions.

• Bioinformatics and Biosystems
• /
• v.2 no.1
• /
• pp.17-23
• /
• 2007

Shot interfering RNA (siRNA) can be used to silence specific gene expression and have many potential therapeutic applications. However, how to design an effective siRNA is still not clear. Highly effective siRNA has sequence-specific properties which are low G/C content, low internal stability at the sense strand 3'-terminus, sense strand base bias(position 1 is G/C, position 19 is /AU). Recently, mRNA secondary structure playsan important role in RNAi. Target site of siRNA in high-ordered structure (i.e hairpin loop, multi loop) or base pair of many hydrogen bonds dramatically reduce function of siRNA mediated gene silencing. Possible off-target effects of siRNA is detecting from BLAST search.

• Applied Biological Chemistry
• /
• v.5
• /
• pp.7-21
• /
• 1964

가잠(家蠶)(Bombyx mori)의 잠체(蠶體), 용체 및 견사선(絹絲腺)(후부(後部))에서 phenol법(法)으로 RNA를 추출(抽出)하여 RNA의 nucleotide 조성(組成)(mole ratio)을 살피는 한편, 견사선(絹絲腺)(후부(後部))에서 초원심법(超遠沈法)으로 r-RNA, s-RNA를 분리(分離)하여 이에 대(對)한 nucleotide조성(組成)을 조사(調査)하고 또 가잠견사선(家蠶絹絲腺)과 비교(比較)할 목적(目的)으로 거미 방적선(紡績腺)의 t-RNA를 분리(分離)하여 nucleotide성분(成分)을 측정(測定)하여 다음과 같은 결과(結果)를 얻었다. 1) 잠란(蠶卵)에 있어서 이것을 마수(磨粹), 탈지(脫脂) 후(後) lysozyme을 작용(作用)시키고 10% NaCl용액(溶液)으로 가열(加熱) 추출(抽出)하는 새방법(方法)을 고찰(考察)하여 RNA의 추출(抽出)이 극난(極難)한 잠란(蠶卵)에서 RNA를 분리(分離)하는데 성공(成功)하였다. 2) 가잠란(家蠶卵), 잠체(蠶體), 용체 및 견사선(絹絲腺)(후부(後部))의 t-RNA nucleotide 조성(組成)은 다음과 같다. 시료(試料) $\frac{G+C}{A+U}$ $\frac{G+U}{A+C}$ $\frac{Pu}{Py}$ 가잠란(家蠶卵)의 RNA 1.14 1.24 0.99 가잠체(家蠶體)의 RNA 1.40 1.36 0.80 용체의 RNA 1.40 1.33 1.35 후부견사선(後部絹絲腺)의 RNA 1.05 1.32 1.15 이로서 잠체(蠶體). 용체 및 견사선(絹絲腺)의 Pu/Py는 각각(各各) 차이(差異)가 있으나 G+U/A+C는 3자간(者間)에 1.3의 거이 동일(同一)한 수치(數値)를 보여주고 있다. G+C/A+U는 잠체(蠶體)와 용체에 있어서 동일(同一)하나 견사선(絹絲腺)의 그것과는 차이(差異)가 있다. 한편 잠란(蠶卵)에 있어서는 Pu/Py, G+C/A+U, G+U/A+C가 각각(各各) 잠체(蠶體), 용체 및 견사선(絹絲腺)에 있어서와 현저(顯著)한 차이(差異)를 보여주고 있다. G+C/A+U가 1.3이나 되는 RNA의 base ratio를 가진 생물(生物)에 관(關)해서는 아직 보고(報告)된 바 없고 다만 본논문(本論文)의 가잠(家蠶)에 관(關)한 RNA와 속편(續編)인 각종(各種) 패류(貝類) RNA의 nucleotide 조성(組成)에서 모두 1.3에 가까운 수치(數値)를 보여주고 있다. 3) 견사선(絹絲腺)(후부(後部)) t-RNA와 거미 방적선(紡績腺)의 t-RNA의 nucleotide molar ratio 및 견사선(絹絲腺)의 r-RNA, s-RNA nucleotide 조성(組成)은 다음과 같다. 재료(材料) $\frac{G+C}{A+U}$ $\frac{G+U}{A+C}$ $\frac{Pu}{Py}$ 가잠견사선(家蠶絹絲腺)(후부(後部)의 t-RNA 1.05 1.32 1.15의 r-RNA 1.12 1.30 1.20의 s-RNA 1.55 1.33 0.65 지주방적선(蜘蛛紡績腺)의 t-RNA 1.35 1.24 1.16 즉(卽) 가잠견사선(家蠶絹絲腺)(후부(後部))과 거미방적선(紡績腺)의 t-RNA nucleotide 조성(組成)은 Pu/Py가 1.15와 1.16으로서 거이 동일(同一)하지만 G+C/A+U, G+U/A+C에 차이(差異)가 있음을 보았다. 한편 가잠견사선(家蠶絹絲腺)(후부(後部)) r-RNA와 s-RNA의 Pu/Py와 G+C/A+U는 현저(顯著)한 차이(差異)가 있고, G+U/A+C에 있어서는 1.3으로서 거이 동일(同一)한 수치(數値)를 보여주고 있다. 4. 이상(以上)과 같이 잠체(蠶體)에 관(關)한 RNA의 nucleotide 조성(組成)은 소위(所謂) GC-type로서, 현재(現在)까지 문헌(文獻)에 보고(報告)된 각종(各種) 생물(生物)의 RNA의 base ratio에 관(關)하여 비교(比較) 검토(檢討)하였으며, RNA의 nucleotide ratio의 차이(差異)의 의의(意義)에 대(對)하여 고찰(考察)하였다.

• Tuberculosis and Respiratory Diseases
• /
• v.56 no.4
• /
• pp.393-404
• /
• 2004

Background : Nucleic acid hybridization has become an essential technique in the development of our understanding of gene structure and function. The quantitative analysis of hybridization has been used in the measurement of genome complexity and gene copy number. The filter hybridization assay is rapid, sensitive and can be used to measure RNAs complementary to any cloned DNA sequence. Methods : The authors assessed the accuracy, linearity, correlation coefficient and specificity of the hybridization depending on the added dose(0, 1, 5, and $10{\mu}g$) of non-specific rat spleen RNA to hybridization of surfactant protein A mRNA. Filter hybridization assays were used to obtain the equation of standard curve and thereby to quantitate the mRNA quantitation. Results : 1. Standard curve equation of filter hybridization assay between counts per minute (X) and spleen RNA input (Y) was Y=0.13X-19.35. Correlation coefficient was 0.98. 2. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) was Y=0.00066X-0.046. Correlation coefficient was 0.99. 3. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $1{\mu}g$ spleen RNA was Y=0.00056X-0.051. Correlation coefficient was 0.99. 4. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $5{\mu}g$ spleen RNA was Y=0.00065X-0.088. Correlation coefficient was 0.99. 5. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $10{\mu}g$ spleen RNA was Y=0.00051X-0.10. Correlation coefficient was 0.99. Conclusions : Comparison of cpm/filter in a linear range allowed accurate and reproducible estimation of surfactant protein A mRNA copy number irrespective of the addition dosage of non-specific rat spleen RNA over the range $0-10{\mu}g$.

• Toxicological Research
• /
• v.15 no.1
• /
• pp.35-38
• /
• 1999

RNA synthesis rate was measured at different time points after UV irradiation in various xeroderma pigmentosum (XP) cells including complementation groups F and G. The RNA synthesis was assayed by measuring 3H-uridine incorporation. In normal cells, recovery of RNA synthesis was initiated at about 6 hr ager UV irradiation and reached to the same level as in unirradiated cells at 24hr after UV irradiation. By contrast, no such recovery was observed in group F,G XP cells.