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

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Functional Nucleotides of U5 LTR Determining Substrate Specificity of Prototype Foamy Virus Integrase

  • Kang, Seung-Yi;Ahn, Dog-Gn;Lee, Chan;Lee, Yong-Sup;Shin, Cha-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1044-1049
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    • 2008
  • In order to study functional nucleotides in prototype foamy virus (PFV) DNA on specific recognition by PFV integrase (IN), we designed chimeric U5 long terminal repeat (LTR) DNA substrates by exchanging comparative sequences between human immunodeficiency virus type-1 (HIV-1) and PFV U5 LTRs, and investigated the 3'-end processing reactivity using HIV-1 and PFV INs, respectively. HIV-1 IN recognized the nucleotides present in the fifth and sixth positions at the 3'-end of the substrates more specifically than any other nucleotides in the viral DNA. However, PFV IN recognized the eighth and ninth nucleotides as distinctively as the fifth and sixth nucleotides in the reactions. In addition, none of the nucleotides present in the twelfth, sixteenth, seventeenth, eighteenth, nineteenth, and twentieth positions were not differentially recognized by HIV-1 and PFV INs, respectively. Therefore, our results suggest that the functional nucleotides that are specifically recognized by its own IN in the PFV U5 LTR are different from those in the HIV-1 U5 LTR in aspects of the positions and nucleotide sequences. Furthermore, it is proposed that the functional nucleotides related to the specific recognition by retroviral INs are present inside ten nucleotides from the 3'-end of the U5 LTR.

A cDNA Clone for the 5' Exon of Chloroplast ATP Synthase Subunit I Gene (atpF) from Broccoli (Brassica oleracea L. var. Italica) and Its Expression Pattern

  • Choo Bong Hong
    • Journal of Plant Biology
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    • v.38 no.2
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    • pp.137-141
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    • 1995
  • We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.

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Effect of Concentration of Ionic Liquids on Resolution of Nucleotides in Reversed-phase Liquid Chromatography

  • Hua, Jin Chun;Polyakova, Yulia;Row, Kyung-Ho
    • Bulletin of the Korean Chemical Society
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    • v.28 no.4
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    • pp.601-606
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    • 2007
  • The chromatographic behaviors of nucleotides (inosine 5'-monophosphate, uridine 5'-monophosphate, guanosine 5'-monophosphate, and thymine monophosphate disodium salts) on a C18 column were studied with different types of ionic liquids (ILs) as additives for the mobile phase in reversed-phase liquid chromatography (RPLC). Three ILs, 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIm][BF4]), 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIm][BF4]), and 1-ethyl-3-methylimidazolium methylsulfate ([EMIm][MS]), were used. Eluents were composed of water and methanol (90/10%, vol) with the addition of 0.5-13.0 mM of ILs. The effects of the concentration of ILs on retention and separation were investigated and discussed. The results showed that the addition of ILs affects the retention and resolution of the tested compounds. Use of 13.0 mM of [BMIm][BF4] as the eluent modifier resulted in a baseline separation of nucleotides without requiring gradient elution. This study demonstrates that ILs can be potentially applied as a mobile phase modifier in RPLC.

Study on the Taste Characteristics of the Chemical Seasoning (MSG) Mixed with the Various Contents of Nucleotides (핵산함유 화학조미료의 맛특성에 관한 연구)

  • 변진원;황인경
    • Korean journal of food and cookery science
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    • v.3 no.1
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    • pp.71-77
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    • 1987
  • This study was to investigate the synergistic taste effect between monosodium glutamate(MSG) and 5'-ribonucleotides consisted of disodium 5'-inosinate (IMP) and 5'-guanylate (GMP) as 1:1 ratio. Solvent was distilled water. Sensory evaluation with 10 panelists was performed by using ratio scaling method (magnitude estimation). The results were as follows: 1) Taste intensities were increased, as nucleotides content to MSG increased. 2) Multiple regression analyses were carried out with the taste intensity data as a function of nucleotides content at three concentrations of seasonings, 0.025%, 0.05% and 0.1%. 3) Predicted taste intensities ($Y_P$) were calculated from the regression equation. Also taste intensity ratios ($Y_{TR}$)-$Y_{TR}$=$Y_P$/taste intensity of MSG only-were calculated. 4) The taste intensity ratios ($Y_{TR}$) at three concentrations of seasonings in the same nucleotides contents showed about the same. Therefore, instead of above regression equations, only one multiple regression equation expressing $Y_P$ of nucleotides seasonings could be determined, as functions of nucleotides content and seasoning concentration.

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Analysis of a Putative DNA Polymerase I gene in Brevibacterium ammoniagenes. (Brevibacterium ammoniagenes의 DNA Polymerase I 유사 유전자의 분석)

  • 오영필;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.105-110
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    • 2002
  • The sequence of 3,221 nucleotides immediately adjacent to rpsA gene encoding 30S ribosomal protein S1 of Brevibacterium ammoniagenes was determined. A putative open reading frame (ORF) of 2,670 nucleotides for a polypeptide of 889 amino acid residues and a TAG stop codon was found, which is located at a distance of 723 nucleotides upstream from rpsA gene with same translational direction. The deduced amino acid sequence of the ORF was found to be highly homologous to the DNA polymerase I of Streptomyces griseus (75.48%), Rhodococcus sp. ATCC 15963 (56.69%), Mycobacterium tuberculosis (55.46%) and Mycobacterium leprae (53.99%). It was suggested that the predicted product of the ORF is a DNA polymerase I with three functional domains. Two domains of 5 → 3 exonuclease and DNA polymerase are highly conserved with other DNA polymerase I, but 3 → 5 exonuclease domain is less conserved.

Analysis of putative promoter sites in Babesia bovis rap-l and B equi ema-l intergenic nucleotides (Babesia bovis rap-1 및 B equi ema-1 intergenic 뉴클레오타이드에서 프로모터로 추정되는 위치 분석)

  • 곽동미
    • Korean Journal of Veterinary Service
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    • v.27 no.1
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    • pp.95-101
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    • 2004
  • Babesia bovis rap-1 and B equi ema-1 intergenic(IG) nucleotides were analyzed and compared for identifying putative promoter sites using computer programs. The reason to initiate this research was to determine if IG nucleotides of Babesia genes that are predicted to be involved in erythrocyte invasion have functions regulating gene transcription and translation, which can be applied to functional gene knockout. Four IG sequences used included BbIG5(B bovis rap-1 5' IG), BblG3(B bovis rap-1 3' IG), BeIG5(B equi ema-1 5' IG) and BeIG3(B equi ema-1 3' IG). BbIG5 contained a putative promoter at nucleotide 197-246 with a predicted TATA-box and a transcription start site. BbIG3 had a putative promoter at nucleotide 270-320 with two predicted TATA-boxes and a transcription start site. BeIG3 had a putative promoter at nucleotide 155-205 with a predicted TATA-box and a transcription start site. Putative promoter sites in these three sequences mentioned above were identified with score cutoff 0.8, which means detection of about 40% recognized promoters with 0.1-0.4% false positives. In contrast, BeIG5 had a putative promoter at nucleotide 163-213 with score cutoff 0.8, but neither TATA-box nor transcription start site were recognized. However, BeIG5 had a putative promoter at nucleotide 388-438 with a predicted TATA-box and a transcription start site when score cutoff was decreased to 0.18, which means detection of about 70% recognized promoters with 2.2-5.3% false positives. These sequences with putative promoters can be tested if they have functions regulating gene transcription and translation.

A Modeling Study of Co-transcriptional Metabolism of hnRNP Using FMR1 Gene

  • Ro-Choi, Tae Suk;Choi, Yong Chun
    • Molecules and Cells
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    • v.23 no.2
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    • pp.228-238
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    • 2007
  • Since molecular structure of hnRNP is not available in foreseeable future, it is best to construct a working model for hnRNP structure. A geometric problem, assembly of $700{\pm}20$ nucleotides with 48 proteins, is visualized by a frame work in which all the proteins participate in primary binding, followed by secondary, tertiary and quaternary binding with neighboring proteins without additional import. Thus, 40S hnRNP contains crown-like secondary structure (48 stemloops) and appearance of 6 petal (octamers) rose-like architectures. The proteins are wrapped by RNA. Co-transcriptional folding for RNP fibril of FMR1 gene can produce 2,571 stem-loops with frequency of 1 stem-loop/15.3 nucleotides and 53 40S hnRNP beaded structure. By spliceosome driven reactions, there occurs removal of 16 separate lariated RNPs, joining 17 separate beaded exonic structures and anchoring EJC on each exon junction. Skipping exon 12 has 5'GU, 3'AG and very compact folding pattern with frequency of 1 stem-loop per 12 nucleotides in short exon length (63 nucleotides). 5' end of exon 12 contains SS (Splicing Silencer) element of UAGGU. In exons 10, 15 and 17 where both regular and alternative splice sites exist, SS (hnRNP A1 binding site) is observed at the regular splicing site. End products are mature FMR-1 mRNP, 4 species of Pri-microRNAs derived from introns 7,9,15 and 3'UTR of exon17, respectively. There may also be some other regulatory RNAs containing ALU/Line elements as well.

Degradation of Nucleotides and Their Related compounds in Sea Foods during processing and Storage -3 . Degradation of Nucleotides and Their Related compounds in Conger-eel Astroconger myriaster Muscle during Drying- (수산식품(水産食品)의 가공(加工) 및 보관중(保藏中)의 핵산(核酸) 관련물질(關聯物質)의 변화(變化)에 관한 연구(硏究) -3. 붕장어 천일건조중(天日乾燥中)의 핵산관련물질(核酸關聯物質)의 변화(變化)-)

  • Lee, Eung-Ho;Han, Bong-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.1 no.1
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    • pp.17-24
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    • 1972
  • In this paper, the degradation of nucleotides and their related compounds in conger eel muscle during sun drying was studied. The results showed that IMP was dominant in fresh conger eel, showing 75.5% of total nucleotides while the contents of ATP, ADP, AMP, inosine and hypoxanthine were low. The nucleotides tended to degrade rapidly during drying, only 6.2% of IMP remained after seven day's sun drying and ATP, ADP and AMP were also entirely disappeared. In consideration of flavor quality, it was consumed that sun drying is not an effective processing method of conger eel, as far as IMP is concerned.

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Activity of virE promoter in Agrobacterium tumefaciens A348 (Agrobacterium tumefaciens A348에서 virE 프로모터의 활성)

  • 음진성
    • Journal of Plant Biology
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    • v.34 no.4
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    • pp.331-339
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    • 1991
  • To elucidate the regulatory mechanism of virE operon from vir regions (virA, virB, virC, virD, virG, virE) of pTiA6 which have been known to be essential for efficient crown gall tumorigenesis in plants, the activity of the truncated virE, promoter was analyzed. pSM358cd, a recombinant plasmid in which virE :: Tn3-HoHo1 (Tn3-promoterless lacZ) was cloned into SalI site of pVK102, was digested with SalI, and virE :: Tn3-HoHo1 was seperated from pVK102. To construct the truncted virE recombinant plasmids (pJS031, pJS051, pJS102, pJS201, pJS301), 5'-end of vireE promoter was deleted with BAL31 and cloned into pVK102 and then transferred into a. tumefaciens A348(pTiA6). According to the activity of the truncated virE promoter in recombinant plasmids, they were classified into two groups, pJS031, pJS051, pJS101 and pJS201 belong to a functional group and pJS301 is a non-functional. The size of deleted nucleotides of pJS201 and pJS301 seemed to be about 130 nucleotides and about 250 nucleotides from 5'-end of virE promoter, respectively. Hence it was thought that the essential site of the virE promoter was located between about 130th nucleotide and 250th nucleotide from 5'-end of the virE promoter.

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Studies on the Occurance of Highly Phosphorylated Nucleotides in the Differentiating Mycelia of Aspergillus niger and Effects of 8-Azaguanine, Cycloheximide on Sporulation (검정곰팡이의 분화에 있어서 고인산뉴클레오티드의 출현 및 8-아자구아닌, 시클로헥시미드의 영향에 관한 연구)

  • Kim, Jong-Hyup;Han, Hee-Jae
    • The Korean Journal of Mycology
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    • v.12 no.4
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    • pp.141-152
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    • 1984
  • Aspergillus niger van Tieghem was cultured by the method of synchronous and submerged culture. Throughout the culture, sporulation was occured. Highly phosphorylated nucleotides in sporulating mycelia were detected to assure whether the eucaryotic Aspergillus niger produce these substances or not during the differentiation. Phosphorylated nucleotides were extracted from the conidiophore, bearing mycelia and spore forming body, these nucleotides were identified by TLC with P.E.I. cellulose. Guanosine tetraphosphate was found in both phialide forming mycelia and spore forming body. The contents of free amino acids were assayed and its level was found to increase at early stage of sporulation. The effects of 8-azaguanine examined, it was found to prevent spore formation and to made abnormal structure. The effects of inosinic monophosphate and guanosine monophosphate on spore formation were examined, spore formation was enhanced by these nucleotides.

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