DNA construct
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Methods in molecular biology ; vol. 170
1st ed. xi, 264 s.
- MeSH
- DNA MeSH
- molekulární biologie MeSH
- referenční knihy MeSH
- Publikační typ
- příručky MeSH
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- NLK Obory
- biologie
[1st ed.] xxiii, 712 s. : il.
- MeSH
- DNA MeSH
- klonování DNA MeSH
- referenční knihy MeSH
- Publikační typ
- příručky MeSH
- učebnice MeSH
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- NLK Obory
- biomedicínské inženýrství
- biologie
... Strains That Overproduce Antibiotics and Other Secondary Metabolites 154 -- Potentials for Recombinant-DNA ... ... DNA Probes Informative for Diagnosis and Identification 232 -- Introduction 232 -- Single Copy DNA Probes ... ... and Their Use 236 -- Methods for Labeling DNA Probes 248 -- Conclusion 251 -- References 252 -- Chapter ... ... Plasmid Stability of Recombinant DNA Microorganisms 316 -- Introduction 316 -- General Properties of ... ... Clinical Development of Drugs and Biologics -- Produced by Recombinant DNA Technology 569 -- Introduction ...
619 s.
- MeSH
- DNA imunologie MeSH
- epitopy genetika imunologie MeSH
- glykoproteiny genetika imunologie MeSH
- imunizace metody MeSH
- lidský herpesvirus 1 imunologie MeSH
- myši inbrední BALB C imunologie virologie MeSH
- plazmidy genetika imunologie MeSH
- vakcína proti viru herpes simplex genetika imunologie MeSH
- vakcinace metody MeSH
- virové geny imunologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
A novel efficient two-step methodology for the construction of base-functionalized DNA is based on direct aqueous cross-coupling reactions of unprotected nucleoside triphosphates followed by polymerase incorporation. Preliminary applications of the modified DNA in electrochemical detection and bioanalysis are outlined.
Covalent DNA interstrand crosslinks are toxic DNA damage lesions that block the replication machinery that can cause a genomic instability. Ubiquitous abasic DNA sites are particularly susceptible to spontaneous cross-linking with a base from the opposite DNA strand. Detection of a crosslink induces the DNA helicase ubiquitination that recruits NEIL3, a DNA glycosylase responsible for the lesion removal. NEIL3 utilizes several zinc finger domains indispensable for its catalytic NEI domain repairing activity. They recruit NEIL3 to the repair site and bind the single-stranded DNA. However, the molecular mechanism underlying their roles in the repair process is unknown. Here, we report the structure of the tandem zinc-finger GRF domain of NEIL3 and reveal the molecular details of its interaction with DNA. Our biochemical data indicate the preferential binding of the GRF domain to the replication fork. In addition, we obtained a structure for the catalytic NEI domain in complex with the DNA reaction intermediate that allowed us to construct and validate a model for the interplay between the NEI and GRF domains in the recognition of an interstrand cross-link. Our results suggest a mechanism for recognition of the DNA replication X-structure by NEIL3, a key step in the interstrand cross-link repair.
We designed and synthesized nucleosides bearing aminophenyl- or aminonaphthyl-3-methoxychromone fluorophores attached at position 5 of cytosine or thymine and converted them to nucleoside triphosphates. The fluorophores showed solvatochromic fluorescence with strong fluorescence at 433-457 nm in non-polar solvents and very weak fluorescence at 567 nm in alcohols. The nucleosides and nucleotides also showed only negligible fluorescence in alcohols or water. The triphosphates were substrates for DNA polymerase in the enzymatic synthesis of modified DNA probes that showed only very weak fluorescence in aqueous buffer but a significant light-up and blue shift were observed when they interacted with proteins (histone H3.1 or p53 for double-stranded DNA probes or single-strand binding protein for single-stranded oligonucleotide probes). Hence, nucleotides have good potential in the construction of DNA sensors for studying protein-DNA interactions. The modified dNTPs were also transported into cells using a cyclodextrin-based transporter but they were not incorporated into the genomic DNA.
Aqueous Sonogashira cross-coupling reactions of 5-iodopyrimidine or 7-iodo-7-deazaadenine nucleosides with bile acid-derived terminal acetylenes linked via an ester or amide tether gave the corresponding bile acid-nucleoside conjugates. Analogous reactions of halogenated nucleoside triphosphates gave directly bile acid-modified dNTPs. Enzymatic incorporation of these modified nucleotides to DNA was successfully performed using Phusion polymerase for primer extension. One of the dNTPs (dCTP bearing cholic acid) was also efficient for PCR amplification.
- MeSH
- denaturace nukleových kyselin MeSH
- DNA-dependentní DNA-polymerasy metabolismus MeSH
- DNA chemie metabolismus MeSH
- nukleosidy chemická syntéza chemie MeSH
- nukleotidy chemická syntéza chemie metabolismus MeSH
- Thermococcaceae enzymologie MeSH
- žlučové kyseliny a soli chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
DNA has played an early and powerful role in the development of bottom-up nanotechnologies, not least because of DNA's precise, predictable, and controllable properties of assembly on the nanometer scale. Watson-Crick complementarity has been used to build complex 2D and 3D architectures and design a number of nanometer-scale systems for molecular computing, transport, motors, and biosensing applications. Most of such devices are built with classical B-DNA helices and involve classical A-T/U and G-C base pairs. However, in addition to the above components underlying the iconic double helix, a number of alternative pairing schemes of nucleobases are known. This review focuses on two of these noncanonical classes of DNA helices: G-quadruplexes and the i-motif. The unique properties of these two classes of DNA helix have been utilized toward some remarkable constructions and applications: G-wires; nanostructures such as DNA origami; reconfigurable structures and nanodevices; the formation and utilization of hemin-utilizing DNAzymes, capable of generating varied outputs from biosensing nanostructures; composite nanostructures made up of DNA as well as inorganic materials; and the construction of nanocarriers that show promise for the therapeutics of diseases.
Recognition and processing by cellular proteins of DNA modified by platinum complexes have been suggested to be relevant to the mechanism of their antitumor activity. Platinum complexes form on DNA various mono- and bifunctional adducts. It has already been described by other authors that intrastrand cross-links formed on DNA by antitumor cis-diamminedichloroplatinum(II) (cisplatin) between neighboring purine residues are recognized by several DNA-binding proteins. In contrast, these proteins do not recognize the intrastrand cross-links formed on DNA by cisplatin or its clinically ineffective trans isomer (transplatin) between nonadjacent base residues. An eventuality heretofore not addressed is that DNA interstrand cross-links (ICLs) of platinum compounds may be recognized by and bound to DNA-binding proteins. DNA probes of 110 base pairs (bp) were constructed containing five equally spaced ICLs of cisplatin or transplatin. These ICLs were formed at specific sites at which these adducts are preferentially formed in natural DNA. Gel electrophoresis mobility shift and competition assays with these probes were used to investigate the specific recognition and binding of the calf thymus HMG1 protein to the DNA ICLs of both platinum isomers. The ICL of antitumor cisplatin was recognized by and bound to the HMG1 protein with a similar affinity as the 1,2-intrastrand d(GpG) cross-link of this drug. The protein binding to the ICL is selective for the DNA modification by cisplatin, but not by chemotherapeutically inactive transplatin.(ABSTRACT TRUNCATED AT 250 WORDS)
- MeSH
- adukty DNA * metabolismus MeSH
- cisplatina * metabolismus MeSH
- DNA footprinting MeSH
- DNA vazebné proteiny * metabolismus MeSH
- endodeoxyribonukleasy metabolismus MeSH
- kompetitivní vazba MeSH
- konformace nukleové kyseliny MeSH
- molekulární sekvence - údaje MeSH
- poškození DNA MeSH
- proteiny s vysokou pohyblivostí metabolismus MeSH
- protinádorové látky * metabolismus MeSH
- reagencia zkříženě vázaná * metabolismus MeSH
- sekvence nukleotidů MeSH
- substrátová specifita MeSH
- vazba proteinů MeSH
- Publikační typ
- práce podpořená grantem MeSH
- srovnávací studie MeSH
