• Title/Summary/Keyword: form I DNA

Search Result 100, Processing Time 0.031 seconds

Benzo(a)pyrene-Triglyceride Adduct: a Potential Molecular Biomarker for Carcinogen Exposure

  • Lee, Byung-Mu
    • Proceedings of the Korean Society of Toxicology Conference
    • /
    • 2002.05b
    • /
    • pp.20-45
    • /
    • 2002
  • In this study, we demonstrated the in vitro and in vivo formation of carcinogen-lipid adduct and its correlation with DNA or protein adducts. The lipids from serum or hepatocyte membranes of Spragu-Dawley rats. human serum, and standard major lipids were in vitro reacted with benzo[a]pyrene(BP) and BP metabolites. 7,8-Dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]-pyrene(BPDE-I), an ultimate carcinogenic form of BP, was covalently bound to triglyceride(TG). BPDE-I-TG adducts isolated by thin-layer chromatography (TLC) were further detected by high performance liquid chromatography(HPLC). TGs, including triolein, tripalmitin and tristearin, showed positive reactions with BPDE-I. However, cholesterol, phospholipids(Phosphatidylcholine, phosphatidyl-ethanolamine, phosphatidyl-inositol and sphingomyelin) and nonesterified fatty acids(palmitic acid, oleic acid, linoleic acid and stearic acid) did not react with BPDE-I. In addition, other BP metabolites (BP-phenols and -diols) did not react with TG, which TG appeared to be the most reactive lipid yet studied with respect to its ability to form an adduct with BPDE-I. There was a clear-cut dose-respect to its ability to form an adduct with BPDE-I-lipid adduct in vitro between TG and [1,3-3H]BPDE-I. In an animal study, BPDE-I-TG was also formed in the serum of rats orally treated with BP(25 mg/rat). Also, obvious correlations between [3H]BP related-biomolecule adducts (DNA, protein) or lipid damage and the BPDE-I-TG adduct were obtained in various tissues of mice i.p. treated with [3H]BP. These data suggest that TG can form an adduct with BPDE-I, as do other macromolecules (DNA, RNA, and protein). Therefore, a carcinogen-lipid adduct would be a useful biomarker for chemical carcinogenesis research and cancer risk assessment.

  • PDF

Cloning and Characterization of cDNA for Korean Rockfish (Sebastes schlegeli ) Insulin-like Growth Factor-I

  • Kwon, Mi-Jin;Jo, Jae-Yoon;Nam, Taek-Jeong
    • Journal of Marine Bioscience and Biotechnology
    • /
    • v.1 no.2
    • /
    • pp.119-125
    • /
    • 2006
  • To understand the comprehensive mechanisms of biological function for insulin-like growth factor-I (IGF-I) in vertebrates, we have investigated the cDNA sequence of this gene in the korean rockfish (Sebastes schlegeli). The mature form of korean rockfish IGF-I was found to be comprised of 67 amino acid residues, showing about a 7 kDa molecular weight. In this study, we used the polymerase chain reaction (PCR) to obtain a korean rockfish IGF-I (KR IGF-I) cDNA fragment, and methods of rapid amplification of cDNA ends (RACE) to obtain a full length of the KR IGF-I sequence. The KR IGF-I encoded for a predicted amino acid sequence showed identities of 93.6 %, 90.7 %, and 85.4 % in comparison with flounder, chinook salmon, and human IGF-I, respectively. To obtain recombinant biologically active polypeptides, korean rockfish B-C-A-D domains were amplified using the PCR, then the isolated cDNA was expressed in the E. coli BL21(DE3). The recombinant KR IGF-I protein biological function was measured by stimulation of [$^3H$] thymidine incorporation, suggesting the cDNA codes for the korean rockfish proIGF-I.

  • PDF

Phylogeny of the Polygonatum odoratum Complex Inferred from Multiple cpDNA and Nuclear RNApol2_i23 Sequence Data (Ruscaceae) (엽록체 DNA 및 핵 DNA RNApol2_i23에 근거한 둥굴레복합체 (Ruscaceae)의 계통 연구)

  • Park, Jeong-Mi;Chung, Kyong-Sook;Oh, Byoung-Un;Jang, Chang-gee
    • Korean Journal of Plant Taxonomy
    • /
    • v.41 no.4
    • /
    • pp.353-360
    • /
    • 2011
  • The sequence data of the plastid DNA (trnL-F IGS, trnL intron, and trnH-psbA) and nuclear DNA (RNApol2_i23) markers were utilized to study phylogenetic relationships among the taxa in the Polygonatum odoratum complex (Ruscaceae). European P. odoratum individuals form a clade with a high bootstrap value, which is a sister to the clade of Korean P. odoratum var. odoratum, P. odoratum var. pluriflorum and P. robustum. The formation of the clade with P. odoratum var. odoratum, P. robustum, and one accession of P. odoratum var. pluriflorum indicates geological speciation in isolated populations in the islands following dispersal events from the mainlands. All data sets form two major clades, which are congruent with the subgroups divided by the basic chromosome numbers (x = 9 and x = 10). Although it is not easy to test the hypothesis of the decrease in the basic chromosome number due to scatter taxon sampling in this study, the molecular data strongly suggested that aneuploidy plays an important role in lineage diversification in the genus Polygonatum. The cytological data was not strongly supported by the cpDNA sequences. Further investigations of the cytological, morphological, and geographical characteristics with comprehensive sampling are desired to understand the evolution and lineage diversification in the genus.

Phage Particle Proteins and Genomic Characterization of the Lactobacillus plantarum Bacteriophage SC 921. (Lactobacillus plantarum Bacteriophage SC 921의 phage particle protein 및 genome의 특성)

  • 김재원;신영재;심영섭;유승구;윤성식
    • Microbiology and Biotechnology Letters
    • /
    • v.26 no.2
    • /
    • pp.117-121
    • /
    • 1998
  • Bacteriophage SC 921 of Lactobacillus plantarum, isolated from kimchi, showed high lytic effects at 0.2 M.O.I. level. The phage particle contained 4 major proteins (48, 34, 32, 29 kDa). Intact DNA of phage SC 921 is a double stranded linear molecule, and the genomic size is approximately 66.5 kilobase pairs (kbp). Restriction analysis of the genome showed that Sma I gave single site cut and Xba I gave 2 site cuts, while Cla I, Kpn I, and EcoR I formed 4, 5, and 6 cuts, respectively. Hind III digested phage DNA to many fragments. A restriction map of genomic DNA was constructed using the restriction endonuclease Kpn I, Sma I, and Xba I. Bacteriophage SC 921 was compared with B2 phage which had been reported to infect Lactobacillus plantarum ATCC 8014(KCCM l1322). Bacteriophage SC 921 differs from B2 phage at least in thr size of its genome and phage particle proteins.

  • PDF

In vitro Evidence that Purified Yeast Rad27 and Dna2 are not Stably Associated with Each Other Suggests that an Additional Protein(s) is Required for a Complex Formation

  • Bae, Sung-Ho;Seo, Yeon-Soo
    • BMB Reports
    • /
    • v.33 no.2
    • /
    • pp.155-161
    • /
    • 2000
  • The saccharomyces cerevisiae Rad27, a structure-specific endonuclease for the okazaski fragment maturation has been known to interact genetically and biochemically with Dna2, an essential enzyme for DNA replication. In an attempt to define the significance of the interaction between the two enzymes, we expressed and purified both Dna2 and Rad27 proteins. In this report, Rad27 could not form a complex with Dna2 in the three different analyses. The analyses included glycerol gradient sedimentation, protein-column chromatography, and coinfection of baculoviruses followed by affinity purification. This is in striking contrast to the previous results that used crude extracts. These results suggest that the interaction between the two proteins is not sufficiently stable or indirect, and thus requires an additional protein(s) in order for Rad27 and Dna2 to form a stable physical complex. This result is consistent with our genetic findings that Schizosaccharomyces pombe Dna2 is capable of interacting with several proteins that include two subunits of polymerase $\delta$, DNA ligase I, as well as Fen-1. In addition, we found that the N-terminal modification of Rad27 abolished its enzymatic activity. Thus, as suspected, we found that on the basis of the structure determination, N-terminal methionine indeed plays an important role in the nucleolytic cleavage reaction.

  • PDF

A Pattern Matching Extended Compression Algorithm for DNA Sequences

  • Murugan., A;Punitha., K
    • International Journal of Computer Science & Network Security
    • /
    • v.21 no.8
    • /
    • pp.196-202
    • /
    • 2021
  • DNA sequencing provides fundamental data in genomics, bioinformatics, biology and many other research areas. With the emergent evolution in DNA sequencing technology, a massive amount of genomic data is produced every day, mainly DNA sequences, craving for more storage and bandwidth. Unfortunately, managing, analyzing and specifically storing these large amounts of data become a major scientific challenge for bioinformatics. Those large volumes of data also require a fast transmission, effective storage, superior functionality and provision of quick access to any record. Data storage costs have a considerable proportion of total cost in the formation and analysis of DNA sequences. In particular, there is a need of highly control of disk storage capacity of DNA sequences but the standard compression techniques unsuccessful to compress these sequences. Several specialized techniques were introduced for this purpose. Therefore, to overcome all these above challenges, lossless compression techniques have become necessary. In this paper, it is described a new DNA compression mechanism of pattern matching extended Compression algorithm that read the input sequence as segments and find the matching pattern and store it in a permanent or temporary table based on number of bases. The remaining unmatched sequence is been converted into the binary form and then it is been grouped into binary bits i.e. of seven bits and gain these bits are been converted into an ASCII form. Finally, the proposed algorithm dynamically calculates the compression ratio. Thus the results show that pattern matching extended Compression algorithm outperforms cutting-edge compressors and proves its efficiency in terms of compression ratio regardless of the file size of the data.

Inhibitory Effect of Red Bean (Phaseolus angularis) Hot Water Extracts on Oxidative DNA and Cell Damage (팥(Phaseolus angularis) 열수 추출물의 산화적 DNA와 세포 손상 억제 효과)

  • Park, Young-Mi;Jeong, Jin-Boo;Seo, Joo-Hee;Lim, Jae-Hwan;Jeong, Hyung-Jin;Seo, Eul-Won
    • Korean Journal of Plant Resources
    • /
    • v.24 no.2
    • /
    • pp.130-138
    • /
    • 2011
  • In this study, we evaluated the protective effects of the hot water extract from red bean (Phaseolus angularis) against oxidative DNA and cell damage induced by hydroxyl radical. The antioxidant activities were evaluated by hydroxyl radical and hydrogen peroxide scavenging assay, and $Fe^{2+}$-chelating assay. Although the extract with hot water didn't scavenge the hydroxyl radical, it removed and chelated hydrogen peroxide and ferrous iron necessary for the induction of hydroxyl radical by 71% and 64% at 200 ${\mu}g/ml$, respectively. Its protective effect on oxidative DNA damage was carried using ${\Psi}$X-174 RF I plasmid DNA comparing the conversion level of supercoiled form of the plasmid DNA into open-circular form and linear form and the expression level of phospho-H2AX in NIH 3T3 cells. In ${\Psi}$X-174 RF I plasmid DNA cleavage assay, it inhibited oxidative DNA damage by 96% at 200 ${\mu}g/ml$. Also, it decreased the expression of phospho-H2AX by 50.1% at 200 ${\mu}g/ml$. Its protective effect against oxidative cell damage was measured by MTT assay and the expression level of p21 protein in NIH 3T3 cells. In MTT assay for the protective effect against the oxidative cell damage, it inhibited the oxidative cell death and the abnormal cell growth induced by hydroxyl radical. Also, it inhibited p21 protein expression by 98% at 200 ${\mu}g/ml$. In conclusion, the results of the present studies indicate that hot water extract from red bean exhibits antioxidant properties and inhibit oxidative DNA damage and the cell death caused by hydroxyl radical.

Suppression of AP-1 Activity by Tanshinone and Cancer Cell Growth Inhibition

  • 박세연;송지성;이덕근;양철학
    • Bulletin of the Korean Chemical Society
    • /
    • v.20 no.8
    • /
    • pp.925-928
    • /
    • 1999
  • The process of transcription is the major point at which gene expression is regulated. The jun and fos families of eukaryotic transcription factor heterodimerize to form complexes capable of binding 5'-TGAGTCA-3'DNA elements (AP-1 binding site). To search for the inhibitors of the jun-fos-DNA complex formation, several natural products extracts were screened and methanol extract of tanshen (the dried roots of Salvia miltiorrhiza Bunge) showed remarkable inhibitory activity. The active compounds of the extracts were purified using re-peated column chromatography and recrystallization. Their structures were identified as tanshinone I and tanshinone IIA. Through the electrophoresis mobility shift assay and cell cytotoxicity test, tanshinone I and tanshinone IIA were identified as inhibitors that suppress not only AP-1 function but also the cell proliferation. Tanshinone I also suppressed the jun-fos-DNA complex formation in TPA-induced NIH 3T3 cells.

Nucleotide Sequence of a Truncated Proteinase Inhibitor I Gene of Potato (감자에서 분리된 절단형 단백질분해효소 억제제 I 유전자의 염기서열)

  • 이종섭
    • Journal of Plant Biology
    • /
    • v.33 no.4
    • /
    • pp.303-307
    • /
    • 1990
  • A genomic clone carrying a proteinase inhibitor I sequence was isolated and characterized. The clone contained a 0.7 kb EcoRI fragment hybridized with tomato inhibitor I cDNA. The nucleotide sequence of the EcoRI fragment revealed presence of a truncated form of a proteinase inhibitor I gene of potato. The truncated gene contained the 5' flanking region and the first exon of a functional proteinase inhibitor I gene. Although the 5' flanking region contained the regulatory sequences TATAAA and CCACT, a deletion of 40 bp occurred between them.

  • PDF

Mitochondrial genome mutations in mesenchymal stem cells derived from human dental induced pluripotent stem cells

  • Park, Jumi;Lee, Yeonmi;Shin, Joosung;Lee, Hyeon-Jeong;Son, Young-Bum;Park, Bong-Wook;Kim, Deokhoon;Rho, Gyu-Jin;Kang, Eunju
    • BMB Reports
    • /
    • v.52 no.12
    • /
    • pp.689-694
    • /
    • 2019
  • Ethical and safety issues have rendered mesenchymal stem cells (MSCs) popular candidates in regenerative medicine, but their therapeutic capacity is lower than that of induced pluripotent stem cells (iPSCs). This study compared original, dental tissue-derived MSCs with re-differentiated MSCs from iPSCs (iPS-MSCs). CD marker expression in iPS-MSCs was similar to original MSCs. iPS-MSCs expressed higher in pluripotent genes, but lower levels in mesodermal genes than MSCs. In addition, iPS-MSCs did not form teratomas. All iPSCs carried mtDNA mutations; some shared with original MSCs and others not previously detected therein. Shared mutations were synonymous, while novel mutations were non-synonymous or located on RNA-encoding genes. iPS-MSCs also harbored mtDNA mutations transmitted from iPSCs. Selected iPS-MSCs displayed lower mitochondrial respiration than original MSCs. In conclusion, screening for mtDNA mutations in iPSC lines for iPS-MSCs can identify mutation-free cell lines for therapeutic applications.