scholarly journals Termination sites of the in vitro nick-translation reaction on DNA that had photoreacted with psoralen.

1983 ◽  
Vol 80 (18) ◽  
pp. 5540-5544 ◽  
Author(s):  
J. G. Piette ◽  
J. E. Hearst
Keyword(s):  
Nick Translation

scholarly journals Escherichia coli β-clamp slows down DNA polymerase I dependent nick translation while accelerating ligation

2018 ◽  
Author(s):  
Amit Bhardwaj ◽  
Debarghya Ghose ◽  
Krishan Gopal Thakur ◽  
Dipak Dutta
Keyword(s):  
Escherichia Coli ◽  
Dna Polymerase ◽  
Nick Translation ◽  
Dna Polymerase I ◽  
Okazaki Fragments ◽  
Pol I ◽  
Dna Strands ◽  
Translation Property ◽  
Polymerase I

AbstractThe nick translation property of DNA polymerase I (Pol I) ensures the maturation of Okazaki fragments by removing primer RNAs and facilitating ligation. However, prolonged nick translation traversing downstream DNA is an energy wasting futile process, as Pol I simultaneously polymerizes and depolymerizes at the nick sites utilizing energy-rich dNTPs. Using an in vitro assay system, we demonstrate that the β-clamp of the Escherichia coli replisome strongly inhibits nick translation on the DNA substrate. To do so, β-clamp inhibits the strand displacement activity of Pol I by interfering with the interaction between the finger subdomain of Pol I and the downstream primer-template junction. Conversely, β-clamp stimulates the 5’ exonuclease property of Pol I to cleave single nucleotides or shorter oligonucleotide flaps. This single nucleotide flap removal at high frequency increases the probability of ligation between the upstream and downstream DNA strands at an early phase, terminating nick translation. Besides β-clamp-mediated ligation helps DNA ligase to seal the nick promptly during the maturation of Okazaki fragments.

Detection of DNA Strand Breaks in HeLa Cells in vitro and in Mouse Sarcoma 180 Cells in vivo Induced by an Alkylating Agent, Carboquone, Using in situ Nick Translation

Oncology ◽  
1990 ◽  
Vol 47 (3) ◽  
pp. 282-286 ◽  
Author(s):  
Yoshihiko Maehara ◽  
Hideaki Anai ◽  
Yoshihisa Sakaguchi ◽  
Tetsuya Kusumoto ◽  
Yasunori Emi ◽  
...  
Keyword(s):  
Alkylating Agent ◽  
Dna Strand Breaks ◽  
Nick Translation ◽  
Strand Breaks ◽  
In Situ Nick Translation ◽  
Mouse Sarcoma ◽  
Dna Strand

scholarly journals Preparation of Labeled DNA, RNA, and Oligonucleotide Probes

2022 ◽  
Vol 2022 (1) ◽  
pp. pdb.top100578
Author(s):  
Michael R. Green ◽  
Joseph Sambrook
Keyword(s):  
Nucleic Acids ◽  
In Vitro Transcription ◽  
Heterogeneous Population ◽  
Oligonucleotide Probes ◽  
Nick Translation ◽  
Chain Reaction ◽  
Random Priming ◽  
Polymerase Chain ◽  
Enzymatic Methods

Labeled nucleic acids and oligonucleotides are typically generated by enzymatic methods such as end-labeling, random priming, nick translation, in vitro transcription, and variations of the polymerase chain reaction (PCR). Some of these methods place the label in specific locations within the nucleic acid (e.g., at the 5′ or 3′ terminus); others generate molecules that are labeled internally at multiple sites. Some methods yield labeled single-stranded products, whereas others generate double-stranded nucleic acids. Finally, some generate probes of defined length, whereas others yield a heterogeneous population of labeled molecules. Options available for generating and detecting labeled nucleic acids, as well as advice on designing oligonucleotides for use as probes, is included here.

Ultrastructural Investigations of the Contractile Filaments of Amoebae

1974 ◽  
Vol 32 ◽  
pp. 188-189
Author(s):  
P.L. Moore
Keyword(s):  
Cytoplasmic Streaming ◽  
Three Dimensional ◽  
Freeze Fracture ◽  
Amoeboid Movement ◽  
Different Types ◽  
Chemical Conditions ◽  
Complete Interpretation

Previous freeze fracture results on the intact giant, amoeba Chaos carolinensis indicated the presence of a fibrillar arrangement of filaments within the cytoplasm. A complete interpretation of the three dimensional ultrastructure of these structures, and their possible role in amoeboid movement was not possible, since comparable results could not be obtained with conventional fixation of intact amoebae. Progress in interpreting the freeze fracture images of amoebae required a more thorough understanding of the different types of filaments present in amoebae, and of the ways in which they could be organized while remaining functional.The recent development of a calcium sensitive, demembranated, amoeboid model of Chaos carolinensis has made it possible to achieve a better understanding of such functional arrangements of amoeboid filaments. In these models the motility of demembranated cytoplasm can be controlled in vitro, and the chemical conditions necessary for contractility, and cytoplasmic streaming can be investigated. It is clear from these studies that “fibrils” exist in amoeboid models, and that they are capable of contracting along their length under conditions similar to those which cause contraction in vertebrate muscles.

In Vitro Induction of Crystals of MT Protein by Vinblastine-Colcemid in Mouse Spermatids

1974 ◽  
Vol 32 ◽  
pp. 132-133
Author(s):  
John J. Wolosewick ◽  
John H. D. Bryan
Keyword(s):  
Endoplasmic Reticulum ◽  
Smooth Endoplasmic Reticulum ◽  
Nuclear Ring ◽  
Rough Er ◽  
Distal Direction ◽  
In Vitro Induction

Early in spermiogenesis the manchette is rapidly assembled in a distal direction from the nuclear-ring-densities. The association of vesicles of smooth endoplasmic reticulum (SER) and the manchette microtubules (MTS) has been reported. In the mouse, osmophilic densities at the distal ends of the manchette are the organizing centers (MTOCS), and are associated with the SER. Rapid MT assembly and the lack of rough ER suggests that there is an existing pool of MT protein. Colcemid potentiates the reaction of vinblastine with tubulin and was used in this investigation to detect this protein.

Induction and Differentiation of Dental Epithelium in Vivo and in Vitro

1982 ◽  
Vol 40 ◽  
pp. 142-145
Author(s):  
E. J. Kollar
Keyword(s):  
Connective Tissue ◽  
Oral Mucosa ◽  
Basal Lamina ◽  
Functional Derivatives ◽  
Specific Source ◽  
Dental Epithelium ◽  
Connective Tissue Stroma

The differentiation and maintenance of many specialized epithelial structures are dependent on the underlying connective tissue stroma and on an intact basal lamina. These requirements are especially stringent in the development and maintenance of the skin and oral mucosa. The keratinization patterns of thin or thick cornified layers as well as the appearance of specialized functional derivatives such as hair and teeth can be correlated with the specific source of stroma which supports these differentiated expressions.

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