RESTRICTION ENDONUCLEASE DIGESTION

Restriction Endonuclease Digestion of pGEM (T) PCR Cloning Vectors Containing Mouse NEIL1, 2 or 3 cDNA Inserts



Restriction Endonuclease Digestion of pGEM (T) PCR Cloning Vectors Containing Mouse NEIL1, 2 or 3 cDNA Inserts

Introduction

Involvement of Nei in DNA repair

Rebuild the process of repair of the genome in the course of duplication of cells and, therefore, to understand how "straightens" from injuries that may occur during the process of copying the DNA strands present in the nucleus: this is the aim of the program Research conducted by Dana Branzei. The copying errors, in fact, are the basis of alterations typical of cancer, and when the repair system is jammed and the defect becomes irreversible, there is an accumulation of chromosome alterations, predisposing to cancer. Identify the genes involved in this process of DNA repair may enable us to study drugs that specifically target tumor cells without damaging the genome of healthy cells (Renault et al, 1997).

To ensure that the gene pool of the cell to be correctly transmitted to daughter cells, the genome must be faithfully duplicated before cell division, through a process known as DNA replication, in which the two strands of DNA are copied. It is not unusual during this process occurring lesions in one strand of DNA. Usually these lesions are immediately repaired, because the cell with extraordinary systems safeguard the genome. One of these is known as homologous recombination: a process allows the damaged DNA strand to deviate the copying on the complementary strand healthy and, therefore, to derive from this the information missing or altered. After the repair pathway is resolved (that is "deleted" all trace of the deviation) and the replication process is brought back to the filament to the cell, thus ensuring the continuation of its normal cell cycle. The sumoylation and 'ubiquitination are two control mechanisms of high importance in order to avoid alterations in the chromosomes, and it is precisely the crucial pivot through which the recombination process is finely regulated and coordinated (Ichihara & Kurosawa 1993, 153-154).

DNA glycosylases

A central mystery in the function or site-specific DNA-binding proteins is the detailed mechanism for rapid location and binding or target sites in DNA. Human oxoguanine DNA glycosylase 1 (hOGG1), for example, must search out rare 8-oxoguanine lesions to preventDefault trans version mutations Arising from oxidative stress. Here we report high-speed imaging or single hOGG1 enzyme molecules diffusing along DNA stretched by shear flow. Salt-concentration-dependent measurements reveal That Occurs Such diffusion axis hOGG1 slides in persistent contact with DNA. At near-physiologic pH and salt concentration, hOGG1 has a subsecond DNA-binding time and slides with a diffusion constant as high as 5 × 10 ^ 6 BP2 / s. Such a value approaches the theoretical upper limit for one-dimensional diffusion and indicates an activation barrier for sliding of only 0.5 kcal / mol (1 kcal = 4.2 kJ). This nearly barrier-less Brownian sliding indicates That DNA glycosylases locate lesion bases by a massively redundant search in All which the enzyme Selectively binds 8-oxoguanine ...