title: ÐÐ»Ð¸ÑÐ½Ð¸Ðµ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¹ Ð¡-Ð±ÐµÐ»ÐºÐ° Ð½Ð° ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¸ Ð¼ÐµÑÐ¸Ð»ÑÑÐ°Ð½ÑÑÐµÑÐ°Ð·Ñ Ð¸ ÑÐ½Ð´Ð¾Ð½ÑÐºÐ»ÐµÐ°Ð·Ñ Ð² ÐºÐ»ÐµÑÐºÐµ
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¼Ð°Ð³Ð¸ÑÑÑÐ°
titleType: Subtitle
title: The effect of C-protein concentration on methyltransferase and restriction endonuclease concentrations
Creators: name: ÐÐ½ÑÐ¾Ð½Ð¾Ð²Ð°, ÐÐ°ÑÑ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: 2020
Subjects: ÑÐ¸ÑÑÐµÐ¼Ð° ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸-Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸
Esp1396I
Ð¡-Ð±ÐµÐ»Ð¾Ðº
ÑÐµÐ³ÑÐ»ÑÑÐ¸Ñ
Ð°ÑÐ°Ð±Ð¸Ð½Ð¾Ð·Ð½ÑÐ¹ Ð¾Ð¿ÐµÑÐ¾Ð½
ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð°Ñ Ð¼Ð¸ÐºÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ñ
restriction-modification system
C-protein
regulation
arabinose operone
fluorescent microscopy

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¸Ð·ÑÑÐµÐ½Ð¸Ñ ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ Ð¸ Ð¿ÑÐ¸ÑÐ¸Ð½ Ð¿ÑÐµÐ¾Ð´Ð¾Ð»ÐµÐ½Ð¸Ñ Ð±Ð°ÐºÑÐµÑÐ¸Ð¾ÑÐ°Ð³Ð¾Ð¼ ÑÐ¸ÑÑÐµÐ¼Ñ ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸-Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Esp1396I. ÐÐ»Ñ Ð¸Ð·ÑÑÐµÐ½Ð¸Ñ Ð¼ÐµÑÐ°Ð½Ð¸Ð·Ð¼Ð° ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ ÑÐ¸ÑÑÐµÐ¼Ñ ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸-Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Esp1396I Ð±ÑÐ»Ð° Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð° Ð¿Ð»Ð°Ð·Ð¼Ð¸Ð´Ð½Ð°Ñ ÑÐ¸ÑÑÐµÐ¼Ð°, ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ°Ñ Ð³ÐµÐ½Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð² Venus Ð¸ mCherry Ð¿Ð¾Ð´ Ð¿ÑÐ¾Ð¼Ð¾ÑÐ¾ÑÐ°Ð¼Ð¸ Ð¼ÐµÑÐ¸Ð»ÑÑÐ°Ð½ÑÑÐµÑÐ°Ð·Ñ Ð¸ ÑÐ½Ð´Ð¾Ð½ÑÐºÐ»ÐµÐ°Ð·Ñ ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸ ÑÐ¾Ð¾ÑÐ²ÐµÑÑÑÐ²ÐµÐ½Ð½Ð¾, Ð° ÑÐ°ÐºÐ¶Ðµ Ð¡-Ð±ÐµÐ»Ð¾Ðº Ð¿Ð¾Ð´ Ð¸Ð½Ð´ÑÑÐ¸ÑÑÐµÐ¼ÑÐ¼ Ð¿ÑÐ¾Ð¼Ð¾ÑÐ¾ÑÐ¾Ð¼. Ð¢Ð°ÐºÐ°Ñ ÑÐ¸ÑÑÐµÐ¼Ð° Ð¿Ð¾Ð·Ð²Ð¾Ð»ÑÐµÑ Ð¸Ð·Ð¼ÐµÐ½ÑÑÑ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ñ Ð¡-Ð±ÐµÐ»ÐºÐ° Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Ð¿ÑÑÐµÐ¼ Ð´Ð¾Ð±Ð°Ð²Ð»ÐµÐ½Ð¸Ñ ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÑ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¹ Ð¸Ð½Ð´ÑÐºÑÐ¾ÑÐ° Ð² Ð¿Ð¸ÑÐ°ÑÐµÐ»ÑÐ½ÑÑ ÑÑÐµÐ´Ñ Ð±Ð°ÐºÑÐµÑÐ¸Ð¹. Ð ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ ÑÑÐ¾Ð³Ð¾ Ð²Ð¾Ð·Ð¼Ð¾Ð¶Ð½Ð¾ Ð½Ð°Ð±Ð»ÑÐ´Ð°ÑÑ Ð·Ð° Ð¸Ð·Ð¼ÐµÐ½ÐµÐ½Ð¸ÐµÐ¼ Ð´Ð¸Ð½Ð°Ð¼Ð¸ÐºÐ¸ ÑÐ¸Ð½ÑÐµÐ·Ð° ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð² Ð² Ð·Ð°Ð²Ð¸ÑÐ¸Ð¼Ð¾ÑÑÐ¸ Ð¾Ñ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¸ Ð¡-Ð±ÐµÐ»ÐºÐ° Ð² ÐºÐ»ÐµÑÐºÐ°Ñ. ÐÐ»Ñ Ð´ÐµÑÐµÐºÑÐ¸Ð¸ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ¸Ð¸ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð»Ð¸ Ð¼ÐµÑÐ¾Ð´Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾Ð¹ Ð¼Ð¸ÐºÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ð¸ Ð¸ ÑÐ¿ÐµÐºÑÑÐ¾ÑÐ»ÑÐ¾ÑÐ¸Ð¼ÐµÑÑÐ¸Ð¸. ÐÐ»Ñ Ð¸Ð·ÑÑÐµÐ½Ð¸Ñ Ð·Ð°ÑÐ¸ÑÐ½ÑÑ ÑÐ²Ð¾Ð¹ÑÑÐ² ÑÐ¸ÑÑÐµÐ¼Ñ Esp1396I, Ð¾Ð½Ð° Ð±ÑÐ»Ð° Ð¿Ð¾Ð¼ÐµÑÐµÐ½Ð° Ð² Ð¿Ð»Ð°Ð·Ð¼Ð¸Ð´Ð½ÑÐ¹ Ð²ÐµÐºÑÐ¾Ñ pBR322, Ðº Ð³ÐµÐ½Ð°Ð¼ Ð¼ÐµÑÐ¸Ð»ÑÑÐ°Ð½ÑÑÐµÑÐ°Ð·Ñ Ð¸ ÑÐ½Ð´Ð¾Ð½ÑÐºÐ»ÐµÐ°Ð·Ñ ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸ Ð±ÑÐ»Ð¸ Ð´Ð¾Ð±Ð°Ð²Ð»ÐµÐ½Ñ Ð³ÐµÐ½Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð². ÐÐ»Ñ Ð¸Ð·ÑÑÐµÐ½Ð¸Ñ Ð¿ÑÐ¸ÑÐ¸Ð½ Ð¿ÑÐµÐ¾Ð´Ð¾Ð»ÐµÐ½Ð¸Ñ Ð·Ð°ÑÐ¸ÑÐ½ÑÑ ÑÐ²Ð¾Ð¹ÑÑÐ² ÑÐ¸ÑÑÐµÐ¼Ñ Ð±Ð°ÐºÑÐµÑÐ¸Ð¾ÑÐ°Ð³Ð¾Ð¼ Ðº ÐºÐ»ÐµÑÐºÐ°Ð¼, ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ¸Ð¼ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾-Ð¼ÐµÑÐµÐ½ÑÑ ÑÐ¸ÑÑÐµÐ¼Ñ ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸-Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Esp1396I, Ð´Ð¾Ð±Ð°Ð²Ð»ÑÐ»Ð¸ Ð±Ð°ÐºÑÐµÑÐ¸Ð¾ÑÐ°Ð³ Î» Ð¸ Ð½Ð°Ð±Ð»ÑÐ´Ð°Ð»Ð¸ Ð·Ð° ÑÐ¾Ð´Ð¾Ð¼ Ð²Ð¸ÑÑÑÐ½Ð¾Ð¹ Ð¸Ð½ÑÐµÐºÑÐ¸Ð¸ Ð² Ð¾Ð´Ð¸Ð½Ð¾ÑÐ½ÑÑ Ð±Ð°ÐºÑÐµÑÐ¸ÑÑ Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾Ð¹ Ð¼Ð¸ÐºÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ð¸. Ð ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð½ÑÑ ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½ÑÐ¾Ð² Ð±ÑÐ»Ð¾ Ð¾Ð±Ð½Ð°ÑÑÐ¶ÐµÐ½Ð¾ Ð¾Ð±ÑÐµÐµ ÑÐ¾Ð³Ð»Ð°ÑÐ¸Ðµ Ñ ÑÐµÐ¾ÑÐµÑÐ¸ÑÐµÑÐºÐ¸Ð¼Ð¸ Ð¿ÑÐµÐ´ÑÑÐ°Ð²Ð»ÐµÐ½Ð¸ÑÐ¼Ð¸ Ð¾ ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ ÑÐ¸ÑÑÐµÐ¼Ñ Esp1396I. ÐÑÐµÐ´Ð¿Ð¾Ð»Ð°Ð³Ð°ÐµÑÑÑ, ÑÑÐ¾ Ð·Ð°ÑÐ¸ÑÐ½ÑÐµ ÑÐ²Ð¾Ð¹ÑÑÐ²Ð° Esp1396I, Ð²Ð¾ Ð¼Ð½Ð¾Ð³Ð¾Ð¼ Ð´Ð¾Ð»Ð¶Ð½Ñ Ð·Ð°Ð²Ð¸ÑÐµÑÑ Ð½Ðµ ÑÐ¾Ð»ÑÐºÐ¾ Ð¾Ñ Ð°Ð±ÑÐ¾Ð»ÑÑÐ½ÑÑ ÐºÐ¾Ð»Ð¸ÑÐµÑÑÐ² Ð±ÐµÐ»ÐºÐ¾Ð² ÑÐ¸ÑÑÐµÐ¼Ñ ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸-Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Ð² ÐºÐ»ÐµÑÐºÐµ, Ð½Ð¾ Ð¸ Ð¾Ñ Ð¸Ñ ÑÐ¾Ð¾ÑÐ½Ð¾ÑÐµÐ½Ð¸Ñ. ÐÐ·ÑÑÐµÐ½Ð¸Ðµ Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾Ð¹ Ð¼Ð¸ÐºÑÐ¾ÑÐºÐ¾Ð¿Ð¸Ð¸ ÑÐ¾Ð¾ÑÐ½Ð¾ÑÐµÐ½Ð¸Ñ Ð¼ÐµÑÐ¸Ð»ÑÑÐ°Ð½ÑÑÐµÑÐ°Ð·Ñ Ð¸ ÑÐ½Ð´Ð¾Ð½ÑÐºÐ»ÐµÐ°Ð·Ñ ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸ Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Ñ ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾-Ð¼ÐµÑÐµÐ½Ð¾Ð¹ ÑÐ¸ÑÑÐµÐ¼Ð¾Ð¹ Esp1396I, Ð¿Ð¾Ð´Ð²ÐµÑÐ³ÑÐ¸ÑÑÑ Ð²Ð¸ÑÑÑÐ½Ð¾Ð¹ Ð¸Ð½ÑÐµÐºÑÐ¸Ð¸, Ð¿Ð¾Ð´ÑÐ²ÐµÑÐ´Ð¸Ð»Ð¾, ÑÑÐ¾ Ð¾ÑÐ½Ð¾ÑÐµÐ½Ð¸Ñ Ð±ÐµÐ»ÐºÐ¾Ð² Ð² ÐºÐ»ÐµÑÐºÐ°Ñ, Ð¿Ð¾Ð³Ð¸Ð±ÑÐ¸Ñ Ð² ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ Ð²Ð¸ÑÑÑÐ½Ð¾Ð¹ Ð¸Ð½ÑÐµÐºÑÐ¸Ð¸, Ð¸ Ð¾ÑÐ½Ð¾ÑÐµÐ½Ð¸Ñ Ð±ÐµÐ»ÐºÐ¾Ð² Ð² Ð½ÐµÐ¸ÑÐ¸ÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ ÐºÐ»ÐµÑÐºÐ°Ñ, ÑÑÐ°ÑÐ¸ÑÑÐ¸ÑÐµÑÐºÐ¸ Ð¾ÑÐ»Ð¸ÑÐ°ÑÑÑÑ.
description: This work is devoted to the investigation of Esp1396I restriction-modification system mechanism of regulation and reasons of its protection overcoming by bacteriophage Î». To study the regulation mechanism of Esp1396I restriction-modification the set of plasmids containing Venus and mCherry fluorescent protein genes under methyltransferase and restriction endonuclease promoters respectively and C-protein gene under inducible promoter were used. Such a system allows to vary C-protein concentration in cells by adding different inducer concentrations to bacteria nutrient medium. As a result, it is possible to observe changes in the fluorescent protein synthesis dynamics which depend on C-protein concentration in the cells. To detect fluorescence we used methods of fluorescent microscopy and fluorescence spectroscopy. To study Esp1396I system protective properties, this restriction-modification system was inserted into pBR322 plasmid vector and methyltransferase and restriction endonuclease genes were fused with Venus and mCherry fluorescent protein genes respectively. To study the reasons of restriction-modification system protection overcoming by bacteriophage viral infection in single cells carrying fluorescently-labeled Esp1396I system was observed by fluorescence microscopy. As a result, general agreement was found with the theoretical view on the Esp1396I restriction-modification system regulation. It was assumed that Esp1396I protective properties should not only depend on absolute restriction-modification system proteins amount, but also their ratio. Study of methyltransferase to restriction endonuclease ratio in single cells by fluorescence microscopy method was implemented in cell containing fluorescently-labeled system Esp1396I. We found that the ratio of restriction-modification proteins in infected and non-infected cells are statistically different.