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Beginning with the Escherichia coli?


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DNA topoisomerases are ubiquitous in nature and have been shown to play critical roles in most p- cesses involving DNA, including DNA replication, transcription, and rec- bination. These enzymes further constitute the cellular targets of a number of clinically important antibacterial and anticancer agents.

Thus, further studies of DNA topology and DNA topoisomerases are critical to advance our und- standing of the basic biological processes required for cell cycle progression, cell division, genomic stability, and development. In addition, these studies will continue to provide critical insights into the cytotoxic action of drugs that target DNA topoisomerases. Such mechanistic studies have already played an important role in the development and clinical application of antimicrobial and chemotherapeutic agents.

The two volumes of DNA Topoisomerase Protocols are designed to help new and established researchers investigate all aspects of DNA topology and the function of these enzymes. The chapters are written by prominent investigators in the field and provide detailed background information and st- by-step experimental protocols. Passar bra ihop.

Recensioner i media. The exploration of the chemical diversity of the naturally occurring lignans and neolignans has resulted in the characterization of many interesting lead compounds in various therapeutic areas; it is thus reasonable to expect that many more leads have yet to be discovered and developed, mainly for antitumor drugs. Jeong-Youn, J. Food Chem. Azarova, A. USA , , Nitiss, J. Drugs , 3 , Oppegard, L. Chen, H. Silva, M. Nova , 26 , Castro-Faria-Neto, H. Castro, M. Serra, M. Verza, M. Morais, S.

DNA topology research group | SBU-Structural Biology Unit

Batista, A. Nova , 33 , Yamaguchi, L. Aguiar, R. It is possible that this is due to the different experimental protocols used in analysis of the cleavage sites [ 25 , 26 ]. Besides M. It remains unclear which part of the type IA enzyme structure determines the cleavage site selectivity. A new procedure for expression and purification of recombinant MtTOP protein in high yield has been described.

The preferred DNA cleavage sites of MtTOP have limited sequence specificity but contain a C nucleotide in the -4 position, similar to most bacterial topoisomerase I and archeal reverse gyrase cleavage sites characterized previously. The recombinant protein in the soluble lysate was allowed to bind to Ni-NTA agarose Qiagen and packed into a column. The DNA was electrophoresed in a 1. The gel was stained with ethidium bromide and photographed over UV light. One unit of enzyme was defined as the least quantity of the enzyme required for complete relaxation of negatively supercoiled DNA under the given reaction conditions.

DNA sequencing reaction products were generated with the same 5' end labeled primer corresponding to that of the substrate used in the cleavage assay and by following the cycle sequencing procedures according to the manufacturer's instructions SequiTherm DNA sequencing Kit, Epicentre.

The sequencing reaction products were electrophoresed next to lanes containing cleavage products to identify the cleavage sites. Annu Rev Biophys Biomol Struct. Nat Rev Mol Cell Biol. Infectious Disorders — Drug Targets.

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Drlica K: Mechanism of fluoroquinolone action. Curr Opin Microbiol. Hooper DC: Bacterial topoisomerases, anti-topoisomerases, and anti-topoisomerase resistance. Clin Infect Dis.

Annu Rev Biochem. Biochim Biophys Acta. Curr Pharm Des. World Health Organization: Tuberculosis fact sheet. J Indian Inst Sci. J Mol Biol.

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Methods Mol Biol. Nucleic Acids Res. J Inorg Biochem. Indian J Biochem Biophys.

J Biol Chem. Eur J Biochem. An enzyme with distinct features. Sikder D, Nagaraja V: Determination of the recognition sequence of Mycobacterium smegmatis topoisomerase I on mycobacterial genomic sequences. A very efficient enzyme that functions independently of zinc binding. Proc Natl Acad Sci. BMC Biochem. Mol Cell.

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Doyle SA: High-throughput cloning for proteomics research. Download references. Correspondence to Yuk-Ching Tse-Dinh. AA carried out the relaxation and DNA cleavage assays, and drafted the manuscript. ND and BC developed and carried out the expression and purification protocol. YT conceived the study, and participated in the design and coordination and helped to draft the manuscript.

All authors read and approved the final manuscript. Reprints and Permissions. Search all BMC articles Search. Abstract Background Mycobacterium tuberculosis DNA topoisomerase I is an attractive target for discovery of novel TB drugs that act by enhancing the accumulation of the topoisomerase-DNA cleavage product.