title: ÐÐ¾ÐºÐ°Ð»Ð¸Ð·Ð°ÑÐ¸Ñ Ð¸ ÑÑÐ½ÐºÑÐ¸Ð¸ RecN - Ð¾Ð´Ð½Ð¾Ð³Ð¾ Ð¸Ð· ÐºÐ»ÑÑÐµÐ²ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð² SOS-Ð¾ÑÐ²ÐµÑÐ°
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Localization and functions of RecN - one of the key proteins of SOS response
Creators: name: Ð Ð¾ÑÐµÐºÑÐ°ÐµÐ²Ð°, ÐÐ¸ÐºÑÐ¾ÑÐ¸Ñ
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2023
Subjects: SOS-Ð¾ÑÐ²ÐµÑ
SMC-Ð±ÐµÐ»ÐºÐ¸
Ð±ÐµÐ»Ð¾Ðº RecN
ÐÐÐ-ÑÐ²ÑÐ·ÑÐ²Ð°ÑÑÐ¸Ð¹ Ð±ÐµÐ»Ð¾Ðº
ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð°Ñ Ð¼Ð¸ÐºÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ñ
Escherichia coli
ÑÐµÐ¿Ð°ÑÐ°ÑÐ¸Ñ ÐÐÐ
SOS-response
SMC-proteins
RecN protein
DNA-binding protein
fluorescence microscopy
DNA repair

description: Â  Â  Â  Â  Ð Ð¿ÐµÑÐ²Ð¾Ð¹ Ð³Ð»Ð°Ð²Ðµ Ð´Ð°Ð½Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÑ Ð¿ÑÐµÐ´ÑÑÐ°Ð²Ð»ÐµÐ½Ð¾ Ð¿Ð¾Ð´ÑÐ¾Ð±Ð½Ð¾Ðµ Ð¾Ð¿Ð¸ÑÐ°Ð½Ð¸Ðµ SOS-Ð¾ÑÐ²ÐµÑÐ° Ð¸ ÑÐ¾Ð»Ñ Ð¸ÑÑÐ»ÐµÐ´ÑÐµÐ¼Ð¾Ð³Ð¾ Ð±ÐµÐ»ÐºÐ° RecN Ð² Ð½ÐµÐ¼, Ð° ÑÐ°ÐºÐ¶Ðµ ÑÐµÐ¾ÑÐ¸Ñ Ð¾ SMC-Ð±ÐµÐ»ÐºÐ°Ñ, ÑÐ°Ðº ÐºÐ°Ðº Ð±ÐµÐ»Ð¾Ðº RecN Ð¾ÑÐ½Ð¾ÑÐ¸ÑÑÑ Ðº Ð´Ð°Ð½Ð½Ð¾Ð¼Ñ ÐºÐ»Ð°ÑÑÑ Ð±ÐµÐ»ÐºÐ¾Ð². ÐÐ¾ Ð²ÑÐ¾ÑÐ¾Ð¹ Ð³Ð»Ð°Ð²Ðµ Ð¿ÑÐµÐ´ÑÑÐ°Ð²Ð»ÐµÐ½Ñ Ð¼Ð°ÑÐµÑÐ¸Ð°Ð»Ñ Ð¸ Ð¼ÐµÑÐ¾Ð´Ñ: ÑÐ±Ð¾ÑÐºÐ° Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¸, ÑÑÐ°Ð½ÑÑÐ¾ÑÐ¼Ð°ÑÐ¸Ñ Ð±Ð°ÐºÑÐµÑÐ¸Ð¹ ÑÐ¾Ð·Ð´Ð°Ð½Ð½Ð¾Ð¹ Ð¿Ð»Ð°Ð·Ð¼Ð¸Ð´Ð¾Ð¹, ÑÐ»ÐµÐºÑÑÐ¾-ÑÐ¾ÑÐµÐ· Ð² Ð¿Ð¾Ð»Ð¸Ð°ÐºÑÐ¸Ð»Ð°Ð¼Ð¸Ð´Ð½Ð¾Ð¼ Ð³ÐµÐ»Ðµ, Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ Ð±ÐµÐ»ÐºÐ° Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ Ð°ÑÑÐ¸Ð½Ð½Ð¾Ð¹ ÑÑÐ¾Ð¼Ð°-ÑÐ¾Ð³ÑÐ°ÑÐ¸Ð¸, Ð°Ð½Ð°Ð»Ð¸Ð· ÑÐ´Ð²Ð¸Ð³Ð° ÑÐ»ÐµÐºÑÑÐ¾ÑÐ¾ÑÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ Ð¿Ð¾Ð´Ð²Ð¸Ð¶Ð½Ð¾ÑÑÐ¸ ÐÐÐ Ð² Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ Ð±ÐµÐ»ÐºÐ°, ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð°Ñ Ð¼Ð¸ÐºÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ñ. Ð ÑÑÐµÑÑÐµÐ¹ Ð³Ð»Ð°Ð²Ðµ Ð¿ÑÐµÐ´ÑÑÐ°Ð²Ð»ÐµÐ½Ñ ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÑ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ. Ð£Ð´Ð°Ð»Ð¾ÑÑ Ð¿Ð¾Ð»Ñ-ÑÐ¸ÑÑ Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¸Ðµ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¸, Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ ÐºÐ¾ÑÐ¾ÑÑÑ Ð±ÑÐ»Ð° Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð° ÑÐºÑ-Ð¿ÑÐµÑÑÐ¸Ñ Ð³ÐµÐ½Ð¾Ð², ÐºÐ¾Ð´Ð¸ÑÑÑÑÐ¸Ñ Ð±ÐµÐ»ÐºÐ¸ RecN Ð¸ RecN::mCherry, Ð¿ÑÐ¾Ð²ÐµÑÑÐ¸ ÑÑÐ¿ÐµÑÐ½Ð¾Ðµ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ Ð±ÐµÐ»ÐºÐ¾Ð². ÐÑÐ»Ð° Ð¾ÑÐµÐ½ÐµÐ½Ð° ÑÐ¿Ð¾ÑÐ¾Ð±Ð½Ð¾ÑÑÑ Ð±ÐµÐ»ÐºÐ° RecN ÑÐ²ÑÐ·ÑÐ²Ð°ÑÑÑÑ Ñ ÐÐÐ Ð¸ ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð° Ð»Ð¾ÐºÐ°Ð»Ð¸Ð·Ð°ÑÐ¸Ñ Ð±ÐµÐ»ÐºÐ° RecN::mCherry Ð² ÐºÐ»ÐµÑÐºÐ°Ñ E. coli Ð² Ð½Ð¾ÑÐ¼Ð°Ð»Ñ-Ð½ÑÑ ÑÑÐ»Ð¾Ð²Ð¸ÑÑ Ð¸ Ð² ÑÐ¾ÑÑÐ¾ÑÐ½Ð¸Ð¸ SOS-Ð¾ÑÐ²ÐµÑÐ°. ÐÐ»Ð°Ð²Ð½ÑÐµ Ð²ÑÐ²Ð¾Ð´Ñ Ð¸Ð· ÑÐ°Ð±Ð¾ÑÑ: 1) Ð±ÐµÐ»Ð¾Ðº RecN ÑÐ²ÑÐ·ÑÐ²Ð°ÐµÑÑÑ Ñ Ð¾Ð´Ð½Ð¾Ð½Ð¸ÑÐµÐ²Ð¾Ð¹ Ð¸ Ð´Ð²ÑÐ½Ð¸ÑÐµÐ²Ð¾Ð¹ ÐÐÐ, Ñ Ð¾Ð´Ð½Ð¾Ð½Ð¸ÑÐµÐ²Ð¾Ð¹ ÐÐÐ ÑÐ²ÑÐ·ÑÐ²Ð°Ð½Ð¸Ðµ Ð»ÑÑÑÐµ, ÑÐ¿Ð¾ÑÐ¾Ð±Ð½Ð¾ÑÑÑ ÑÐ²ÑÐ·ÑÐ²Ð°Ð½Ð¸Ñ Ð¿Ð¾Ð²ÑÑÐ°ÐµÑÑÑ Ð¿ÑÐ¸ Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ ÐÐ¢Ð¤ Ð¸ ÑÐ²ÐµÐ»Ð¸ÑÐµÐ½Ð¸Ð¸ ÐµÐ³Ð¾ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¸. 2) Ð² Ð½Ð¾ÑÐ¼Ð°Ð»ÑÐ½ÑÑ ÑÑÐ»Ð¾Ð²Ð¸ÑÑ Ð»Ð¾ÐºÐ°Ð»Ð¸Ð·Ð°ÑÐ¸Ñ ÐÐÐ Ð¸ Ð±ÐµÐ»ÐºÐ° RecN Ð²Ð½ÑÑÑÐ¸ ÐºÐ»ÐµÑÐºÐ¸ Ð½Ðµ ÑÐ¾Ð²Ð¿Ð°Ð´Ð°ÐµÑ, Ð±ÐµÐ»Ð¾Ðº Ð»Ð¾ÐºÐ°Ð»Ð¸Ð·ÑÐµÑÑÑ Ð½Ð° Ð¿Ð¾Ð»ÑÑÐ°Ñ ÐºÐ»ÐµÑÐºÐ¸, Ð²ÑÑÑÐ¸Ð»Ð°ÐµÑ Ð²Ð½ÑÑÑÐµÐ½Ð½ÑÑ Ð¼ÐµÐ¼Ð±ÑÐ°Ð½Ñ, ÑÑÐ¾ Ð¿Ð¾Ð·Ð²Ð¾Ð»Ð¸Ð»Ð¾ ÑÐ´ÐµÐ»Ð°ÑÑ Ð¿ÑÐµÐ´Ð¿Ð¾Ð»Ð¾Ð¶ÐµÐ½Ð¸Ðµ Ð¾ ÑÐ¾Ð¼, ÑÑÐ¾ Ð±ÐµÐ»Ð¾Ðº RecN â Ð°ÑÑÐ¾ÑÐ¸Ð¸ÑÐ¾Ð²Ð°Ð½ Ñ Ð¼ÐµÐ¼Ð±ÑÐ°Ð½Ð¾Ð¹. ÐÐ¾ Ð²ÑÐµÐ¼Ñ SOS-Ð¾ÑÐ²ÐµÑÐ° Ð±ÐµÐ»Ð¾Ðº RecN Ð¿ÑÐµÐ´Ð¿Ð¾Ð»Ð¾Ð¶Ð¸ÑÐµÐ»ÑÐ½Ð¾ Ð¿ÑÐ¸Ð²Ð»ÐµÐºÐ°ÐµÑÑÑ Ðº Ð¼ÐµÑÑÐ°Ð¼ Ð¿Ð¾Ð²ÑÐµÐ¶Ð´ÐµÐ½Ð¸Ñ ÐÐÐ, Ð³Ð´Ðµ, Ð¿Ð¾-Ð²Ð¸Ð´Ð¸Ð¼Ð¾Ð¼Ñ, ÑÑÐ°ÑÑÐ²ÑÐµÑ Ð² Ð¿ÑÐ¾ÑÐµÑÑÐµ ÑÐµÐ¿Ð°ÑÐ°ÑÐ¸Ð¸ ÑÐ¾Ð²Ð¼ÐµÑÑÐ½Ð¾ Ñ Ð±ÐµÐ»ÐºÐ¾Ð¼ RecA.
description: The first chapter of this work provides a detailed description of the SOS-response and the role of RecN protein in it as well as the theory of SMC-proteins as RecN protein belongs to this class of proteins. The second chapter describes materials and methods used in the experiment: assembly of genetic construction, transformation of bacteria with plasmid, electrophoresis in polyacrylamide gel, protein purification by affinity chromatography, analysis of DNA electrophoretic mobility shift in the presence of protein, fluorescent microscopy. The third chapter presents the results of the study. Genetic constructs were obtained to express the genes encoding RecN and RecN::mCherry proteins and to carry out successful protein purification. The ability of RecN protein to bind to DNA was evaluated, and the localization of RecN::mCherry protein in E. coli cells under normal conditions and in SOS-responsive condition was determined. Major conclusions from the work: 1) protein RecN binds to single- and dou-ble-stranded DNA, with single-stranded DNA binding is better, the binding capacity increases in the presence of ATP and increasing of its concentration. 2) under normal conditions the localization of DNA and protein RecN inside the cell does not coincide, the protein is localized at the cell poles, it lines the inner membrane, which suggests that the RecN protein is associated with the membrane. During the SOS response, RecN protein is presumably attracted to DNA damage sites where it appears to participate in the repair process together with RecA protein.