• Title/Summary/Keyword: 5$^1$-nucleotides

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DEGRADATION OF NUCLEOTIDES IN THE MUSCLE OF SEA MUSSEL DURING DRYING (진주담치 건조중의 Nucleotides의 변화)

  • PARK Yeung-Ho;PARK Hwa-Sool;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.163-168
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    • 1974
  • The present paper deals with the degradation of nucleotides in the muscle of sea mussel, Mytilus edulis, during drying. Three kinds of samples, raw, hot-air dried, and steamed-and-hot air dried were prepared and the contents of nucleotides were determined by ion exchange chromatography on columns of Dowex 1, X8. ATP and ADP were dominant in the raw muscle showed about $8{\mu}moles/g$, dry basis, respectively. The rate of degadation of ATP was very slow during drying compared with those of fish. The accumulation of ADP and AMP were observed during drying and the amount of total nucleotides (ATP+ADP+AMP) were not decreased remarkably by drying process. IMP was not detected in the all of the samples examined, however, the contents of inosine and hypoxanthine were increased during drying. In case of inosine contents, the hot-air dried sample marked an exceedingly high value equivalent to 8 times of the raw sample whereas steamed-and-hot air dried sample showed 2 times of raw samples.

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Putative Secondary Structure of Human Hepatitis B Viral X mRNA

  • Kim, Ha-Dong;Choi, Yoon-Chul;Lee, Bum-Yong;Junn, Eun-Sung;Ahn, Jeong-Keun;Kang, Chang-Won;Park, In-Won
    • BMB Reports
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    • v.28 no.6
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    • pp.509-514
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    • 1995
  • A putative secondary structure of the mRNA for the human hepatitis B virus (HBV) X gene is proposed based on not only chemical and enzymatic determination of its single- and double-stranded regions but also selection by the computer program MFOLD for energy minimum conformation under the constraints that the experimentally determined nucleotides were forced or prohibited to base pair. An RNA of 536 nucleotides including the 461-nucleotide HBV X mRNA sequence was synthesized in vitro by the phage T7 RNA polymerase transcription. The thermally renatured transcripts were subjected to chemical modifications with dimethylsulfate and kethoxal and enzymatic hydrolysis with single strand-specific RNase T1 and double strand-specific RNase V1, separately. The sites of modification and cleavage were detected by reverse transcriptase extension of 4 different primers. Many nucleotides could be assigned with high confidence, twenty in double-stranded and thirty-seven in Single-stranded regions. These nucleotides were forced and prohibited, respectively, to base pair in running the recursive RNA folding program MFOLD. The results suggest that 6 different regions (5 within X mRNA) of 14~23 nucleotides are Single-stranded. This putative structure provides a good working model and suggests potential target sites for antisense and ribozyme inhibitors and hybridization probes for the HBV X mRNA.

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Analysis of Nucleotides and Their Derivatives in Renal Tissue of Rat during Ischemia by HPLC (흰쥐의 신허혈에서 HPLC를 이용한 핵산대사산물의 분석)

  • Kim, Seong-Yong;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.9 no.1
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    • pp.90-101
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    • 1992
  • In rat kidney, the changes in concentrations of nucleotides and their derivatives during ischemia induced by renal artery ligation was measured quantitatively with high performance liquid chromatography(HPLC). After the ligation of renal artery for 60minutes, the concentrations of the nucleotides and derivatives were measured. In ischemic tissue, IDP was significantly decreased from $217.4{\pm}12.68{\mu}g$ in control to $80.7{\pm}18.39{\mu}g$ (p<0.01) ; ATP, $307.2{\pm}56.63{\mu}g$ to $47.6{\pm}5.95{\mu}g$ (p<0.01) ; ADP+AMP, $227.1{\pm}7.98{\mu}g$ to $61.4{\pm}3.92{\mu}g$(P<0.01); $NAD^+$, $217.9{\pm}4.49{\mu}g$ to $126.6{\pm}10.44{\mu}g$(P<0.01) ; GTP, $202.5{\pm}23.76{\mu}g$ to $117.7{\pm}14.24{\mu}g$ (P<0.05) ; GMP, $54.5{\pm}9.03{\mu}g$ to $23.7{\pm}0.46{\mu}g$(p<0.05), and inosine, $16.6{\pm}3.45{\mu}g$ to $7.8{\pm}0.87{\mu}g$ (P<0.05). But hypoxanthine and xanthine were significantly increased from $113.0{\pm}15.58{\mu}g$ to $159.7{\pm}12.07{\mu}g$ (P<0.05) and from $87.7{\pm}6.77{\mu}g$ to $173.1{\pm}12.52{\mu}g$ (P<0.01). In ischemic kidney, concentration of ATP was decreased to 39.9% of control at 10 minutes, 19.8% at 30 minutes, and 15.5% at 60 minutes, and ADP+AMP were decreased to 70.3% of control at 10 minutes, 67.3% at 30 minutes, and to 27.0% at 60 minutes, but hypoxanthine and xanthine were increased to 121.5% and 127.1% at 10 minutes, 126.0% and 174.4% at 30 minutes, and 141.4% and 197.3% at 60 minutes Total adenosine nucleotides were decreased to 20.3% of control during 60 minutes of ischemia, but hypoxanthine and xathine were increased to 157.5 % of control. These results suggest that the changes in the concentration of nucleotides and their metabolic derivatives are useful indices of the extents of tissue ischemia in rat kidney.

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Differentially Expressed Genes of Potentially Allelopathic Rice in Response against Barnyardgrass

  • Junaedi, Ahmad;Jung, Woo-Suk;Chung, Ill-Min;Kim, Kwang-Ho
    • Journal of Crop Science and Biotechnology
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    • v.10 no.4
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    • pp.231-236
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    • 2007
  • Differentially expressed genes(DEG) were identified in a rice variety, Sathi, an indica type showing high allelopathic potential against barnyardgrass(Echinochloa crus-galli(L.) Beauv. var. frumentaceae). Rice plants were grown with and without barnyardgrass and total RNA was extracted from rice leaves at 45 days after seeding. DEG full-screening was performed by $GeneFishing^{TM}$ method. The differentially expressed bands were re-amplified and sequenced, then analyzed by Basic Local Alignment Search Tool(BLAST) searching for homology sequence identification. Gel electrophoresis showed nine possible genes associated with allelopathic potential in Sathi, six genes(namely DEG-1, 4, 5, 7, 8, and 9) showed higher expression, and three genes(DEG-2, 3 and 6) showed lower expression as compared to the control. cDNA sequence analysis showed that DEG-7 and DEG-9 had the same sequence. From RT PCR results, DEG-6 and DEG-7 were considered as true DEG, whereas DEG-1, 2, 3, 4, 5, and 8 were considered as putative DEG. Results from blast-n and blast-x search suggested that DEG-1 is homologous to a gene for S-adenosylmethionine synthetase, DEG-2 is homologous to a chloroplast gene for ribulose 1,5-bisphosphate carboxylase large subunit, DEG-8 is homologous to oxysterol-binding protein with an 85.7% sequence similarity, DEG-5 is homologous to histone 2B protein with a 47.9% sequence similarity, DEG-6 is homologous to nicotineamine aminotransferase with a 33.1% sequence similarity, DEG-3 has 98.8% similarity with nucleotides sequence that has 33.1% similarity with oxygen evolving complex protein in photosystem II, DEG-7 is homologous to nucleotides sequence that may relate with putative serin/threonine protein kinase and putative transposable element, and DEG-4 has 98.8% similarity with nucleotides sequence for an unknown protein.

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Nucleotide Analysis in Korean Dairy Products Using High- Performance Liquid Chromatography with Diode Array Detector

  • Won, Jong-Eun;Bang, Han-Yeol;Kwak, Byung-Man;Park, Jong-Su;Kim, Gui-Ran;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.39 no.1
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    • pp.93-101
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    • 2019
  • Nucleotides play important roles in numerous intracellular biochemical processes and are used in infant formulas and other dairy products. However, domestic analytical methods for assessing nucleotide content in products have not yet been established, and therefore, methods for determining nucleotide content are urgently required. A rapid and simple analytical method for determining the content of five types of nucleotides in dairy products was improved using solid phase extraction clean-up and high-performance liquid chromatography with diode array detector. The extraction solvent used in the AOAC method was not well dissolved and was changed to hydrophilic EDTA-Na. In addition, the results obtained using the isocratic elution method and a single wavelength were similar to those obtained using the AOAC method, and the time taken for analysis was shortened from 40 min to 25 min. The process of method validation revealed the following parameters: accuracy (84.69%-102.72%), precision (1.51%-6.82%), linearity (0.999), and limit of detection (cytidine 5'-monophosphate, 0.09 mg/L; uridine 5'-monophosphate, 0.11 mg/L; adenosine 5'-monophosphate, 0.12 mg/L; guanosine 5'-monophosphate, 0.11 mg/L; and inosine 5'-monophosphate, 0.14 mg/L). The method was also used to determine the nucleotide concentration in 25 samples (infant formulas, 1.99-29.39 mg/100 g; and cow milk, 0.28-0.83 mg/100 g). The newly improved method was appropriate for analyzing nucleotides in infant formulas and other dairy products faster when compared to conventional methods.

Quality characteristics of enzymatic anchovy hydrolysates (멸치 효소 가수분해물의 품질특성)

  • Kang, C.S.;Jeong, D.S.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.14 no.1
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    • pp.93-105
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    • 2012
  • This study was carried out to prepare anchovy hydrolysates by using Alcalase, Neutrase and Protamex, and to investigate its quality characteristics. The major free amino acids were glutamic acid and lysine. The total amino acid content of anchovy hydrolysate were glutamic acid(12.6%), lysine(8.56%), valine(7.06%), aspartic acid(5.73%). Major nucleotides content of anchov hydrolysate were 5'-IMP (22.462 mg/100g), 5'-GMP (18.674 mg/100g) and 5'-UMP (1.25 mg/100g). Histamine content of anchovy hydrolysate was 18.1 mg/100g. These results suggested that anchovy hydrolysate could be used as a sauce of basic seasoning.

Sequencing of cDNA Clones Expressed in Adipose Tissues of Korean Cattle

  • Bong, J.J.;Tong, K.;Cho, K.K.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.483-489
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    • 2005
  • To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

DEGRADATION OF ACID SOLUBLE NUCLEOTIDES AND THEIR RELATED COMPOUNDS IN SEA FOODS DURING PROCESSING AND STORAGE 1. Changes of Nucleotides during Drying Process of the Anchovy, Engraulis japonica (수산식품의 가공 및 보장중의 핵산관련물질의 변화에 관한 연구 1. 마른 멸치 제조과정중의 핵산관련물질의 변화)

  • LEE Eung-Ho;PARK Young-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.4 no.1
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    • pp.31-41
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    • 1971
  • The present study was directed to define the degradation pattern of the nucleotides and their related compounds in the muscle of anchovy during drying. Three kinds of samples, fresh, sun dried and boiled-and-dried anchovy, were prepared and the contents of nucleotides and related compounds of samples were determined by ion exchange chromatography. The results obtained are summarized as follows: Almost all of ATP disappeared in both muscle of sun dried and boiled-and-dried anchovy, although the initial content of ATP in fresh muscle was very low ($1.8{\mu}moles/g$, dry basis). But the remainning amount of ADP was considerably high while the other nucleotide almost entirely disappeared. This suggested that the residual ADP is responsible to the 'bound nucleotide' of myofibrils. In general, AMP content was comparatively lower than that of other nucleotides. Among three samples, the boiled-and-dried sample showed relatively higher AMP value than others. The amount of IMP remained in muscle remarkably varied between the boiled-and-dried anchovy and sun dried anchovy, the former's value being sixteen times higher than that of latter. In the contents of inosine and hypoxanthine, the sun dried anchovy marked an exceedingly high value equivalent to 2.7 times of the boiled-and-dried anchovy. In comparison of the ratio of inosine and hypoxanthine, hypoxanthine was accumulating in boiled-and-dried anchovy whereas inosine was in the sun-dried anchovy. Eighty three percent of total nucleotides in the fresh anchovy retained in the boiled-and-dried anchovy and IMP ratio in total nucleotides was $73\%$. On the contrary, the sun dried anchovy showed barely $10\%$ of retention rate and IMP ratio was only $38\%$. Considered from the flavor quality of dried anchovy, so far as concerned IMP content, it may be said that the boiled-and-drying method is more favorable process for dried product of anchovy than the sun-drying method.

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Changes of Nucleotides and their Related Compounds in Cultured and Wild Red Sea Bream and Flounder muscle (양식 및.자연산 도미와 넙치 어육 중의 핵산관련물질의 변화)

  • 이경희;이영순
    • Korean journal of food and cookery science
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    • v.17 no.5
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    • pp.517-522
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    • 2001
  • Changes of nucleotides and their related compounds in raw, cooked and frozen fish muscle were studied with HPLC. Red sea bream(cultured and wild) and flounder(cultured, cultured with Obosan(equation omitted) and wild) were used for this study. In nucleotides, contents of ATP was similar to that of IMP and some of H$\times$R(inosine) and H$\times$(hypoxanthine) were existed in fresh muscle. ATP was decomposed rapidly and contents of IMP became different between cultured and wild fish after 6 hours. The content of IMP was lower in the cultured red sea bream(3.39$\mu$ mole/g) and flounder(3.17$\mu$ mole/g) than in the wi1d red sea bream(7.31$\mu$ mole/g) and flounder(5.03$\mu$ mole/g). But, the flounder cultured with Obosan contained the largest amounts of IMP After 24 hours, K values of cultured fish muscle(27.7%, 28.2%) were higher than that of wild ones(22.8%, 24.3%). The K value of cultured flounder fed with 0.3% Obosan(equation omitted)(25.7%) was between cultured and wild flounder. IMP was the one which existed the most in cooked and frozen muscle. Amounts of H$\times$R and H$\times$ were more in cooked and frozen muscle. than in raw muscle. From these results, we could suggest that the wild one was more palatable and fresher than the cultured one and the palatability of cultured one seemed to be improved depanding on the feed.

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Molecular Cloning of the 3'-Terminal Region of Garlic Potyviruses and Immunological Detection of Their Coat Proteins

  • Song, Sang-Ik;Song, Jong-Tae;Chang, Moo-Ung;Lee, Jong-Seob;Park, Yang-Do
    • The Plant Pathology Journal
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    • v.15 no.5
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    • pp.270-279
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    • 1999
  • cDNAs complementary to the 3'-terminal regions of two potyvirus genomes were cloned and sequenced. The clone G7 contains one open reading frame (ORF) of 1,338 nucleotides and a 3' untranslated region (3'-UTR) of 403 nucleotides at the 3'-end excluding the 3'end poly(A) tail. The putative viral coat protein (CP) shows 55%-92% amino acid sequence homology to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.0 kb by Northern blot analysis. Five cDNA clones were screened out using GPV2 oligonucleotide as a probe. One of these clones, DEA72, which has a longest cDNA insert, contains one ORF of 1,459 nucleotides and a 3'-UTR of 590 nucleotides at the 3'-end excluding the 3'-end poly(A) tail. The putative viral CP shows 57%-88% amino acid sequence homologies to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.6 kb by Northern blot analysis. The results of immunoblot and Northern blot analyses suggest that almost all of the tested garlic plants showing mosaic or streak symptoms are infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus in variable degrees but rarely infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus. Immunoelectron microscopy using anti-DEA72 CP antibody shows that this potyvirus is about 750 nm long and flexuous rod shaped.

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