title: Ð­ÐºÑÐ¿ÑÐµÑÑÐ¸Ñ Ð¸ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ ÐÐÐ-ÑÐ²ÑÐ·ÑÐ²Ð°ÑÑÐµÐ³Ð¾ Ð±ÐµÐ»ÐºÐ° DprA
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: Expression and isolation of DNA-binding protein DprA
Creators: name: Ð¡ÑÐ°Ð½ÐºÐµÐ²Ð¸Ñ, ÐÑÑÑÐ¼
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: 2022
Subjects: ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ñ
Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ
ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°ÑÐ¸Ñ
Ð½Ð°ÑÑÑÐ°Ð»ÑÐ½Ð°Ñ ÑÑÐ°Ð½ÑÑÐ¾ÑÐ¼Ð°ÑÐ¸Ñ
Ð´Ð½Ðº-ÑÐ²ÑÐ·ÑÐ²Ð°ÑÑÐ¸Ðµ Ð±ÐµÐ»ÐºÐ¸
expression
isolation
recombination
natural transformation
dna-binding proteins

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ Ð¸ Ð¾ÑÐ¸ÑÑÐºÐµ ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð½ÑÐ½Ð¾Ð³Ð¾ ÐÐÐ-ÑÐ²ÑÐ·ÑÐ²Ð°ÑÑÐµÐ³Ð¾ Ð±ÐµÐ»ÐºÐ° DprA Ð¸Ð· ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÑ Ð¼Ð¸ÐºÑÐ¾Ð¾ÑÐ³Ð°Ð½Ð¸Ð·Ð¼Ð¾Ð². ÐÐ°Ð´Ð°ÑÐ¸, ÑÐµÑÐ°ÐµÐ¼ÑÐµ Ð² ÑÐ¾Ð´Ðµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ: 1. ÐÐ¿ÑÐ¸Ð¼Ð¸Ð·Ð¸ÑÐ¾Ð²Ð°ÑÑ ÑÑÐ»Ð¾Ð²Ð¸Ñ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ Ð³ÐµÐ½Ð° Ð±ÐµÐ»ÐºÐ° DprA Ð¸Ð· Ð¾ÑÐ³Ð°Ð½Ð¸Ð·Ð¼Ð° Escherichia coli ÑÐ»Ð¸ÑÐ¾Ð³Ð¾ Ñ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾ÑÑÑÑ, ÐºÐ¾Ð´Ð¸ÑÑÑÑÐµÐ¹ 6 Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½ÑÑ Ð³Ð¸ÑÑÐ¸Ð´Ð¸Ð½Ð¾Ð², Ð² ÐºÐ»ÐµÑÐºÐ°Ñ BL21(DE3). 2. ÐÑÑÐ°Ð±Ð¾ÑÐºÐ° ÑÑÐµÐ¼Ñ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ñ Ð±ÐµÐ»ÐºÐ° 6xHis-DprA (E. c.) Ð¼ÐµÑÐ¾Ð´Ð°Ð¼Ð¸ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ð¸. 3. ÐÑÑÐ°Ð±Ð¾ÑÐºÐ° ÑÑÐ»Ð¾Ð²Ð¸Ð¹ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ Ð³ÐµÐ½Ð° Ð±ÐµÐ»ÐºÐ° DprA Ð¸Ð· Ð¾ÑÐ³Ð°Ð½Ð¸Ð·Ð¼Ð° Bacillus subtilis ÑÐ»Ð¸ÑÐ¾Ð³Ð¾ Ñ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾ÑÑÑÑ, ÐºÐ¾Ð´Ð¸ÑÑÑÑÐµÐ¹ 6 Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½ÑÑ Ð³Ð¸ÑÑÐ¸Ð´Ð¸Ð½Ð¾Ð², Ð² ÐºÐ»ÐµÑÐºÐ°Ñ BL21(DE3)pLysE. 4. ÐÑÑÐ°Ð±Ð¾ÑÐºÐ° ÑÑÐµÐ¼Ñ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ñ Ð±ÐµÐ»ÐºÐ° 6xHis-DprA (B. s.) Ð¼ÐµÑÐ¾Ð´Ð°Ð¼Ð¸ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ð¸. ÐÐ±ÑÐµÐºÑ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ â ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð½ÑÐ½ÑÐ¹ Ð±ÐµÐ»Ð¾Ðº DprA. Ð Ð°Ð±Ð¾ÑÐ° Ð²ÑÐ¿Ð¾Ð»Ð½ÐµÐ½Ð° Ð½Ð° Ð±Ð°Ð·Ðµ Ð»Ð°Ð±Ð¾ÑÐ°ÑÐ¾ÑÐ¸Ð¸ Ð½Ð°ÑÑÐ½Ð¾-Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÑÐºÐ¾Ð³Ð¾ ÐºÐ¾Ð¼Ð¿Ð»ÐµÐºÑÐ° Â«ÐÐ°Ð½Ð¾Ð±Ð¸Ð¾ÑÐµÑÐ½Ð¾Ð»Ð¾Ð³Ð¸Ð¸Â» (ÐÐÐ Â«ÐÐ°Ð½Ð¾ÐÐ¸Ð¾Â») Ð² ÑÐ¾ÑÑÐ°Ð²Ðµ Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¾Ð³Ð¾ Ð³Ð¾ÑÑÐ´Ð°ÑÑÑÐ²ÐµÐ½Ð½Ð¾Ð³Ð¾ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¾Ð³Ð¾ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑÐ° ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾. Ð ÑÐ°Ð±Ð¾ÑÐµ Ð±ÑÐ»Ð¸ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ñ Ð±Ð°Ð·Ð¾Ð²ÑÐµ Ð¼Ð¸ÐºÑÐ¾Ð±Ð¸Ð¾Ð»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¸Ðµ Ð¼ÐµÑÐ¾Ð´Ñ, Ð¼ÐµÑÐ¾Ð´Ñ Ð¼Ð¾Ð»ÐµÐºÑÐ»ÑÑÐ½Ð¾Ð¹ Ð±Ð¸Ð¾Ð»Ð¾Ð³Ð¸Ð¸, ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸ÑÐµÑÐºÐ¸Ðµ Ð¼ÐµÑÐ¾Ð´Ñ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ñ ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð½ÑÐ½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð², Ð° ÑÐ°ÐºÐ¶Ðµ ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÐµ Ð¼ÐµÑÐ¾Ð´Ñ ÐºÐ¾Ð½ÑÑÐ¾Ð»Ñ Ð¸ Ð°Ð½Ð°Ð»Ð¸Ð·Ð°. Ð ÑÐ¾Ð´Ðµ Ð²ÑÐ¿Ð¾Ð»Ð½ÐµÐ½Ð¸Ñ ÐÐÐ  Ð±ÑÐ»Ð° Ð´Ð¾ÑÑÐ¸Ð³Ð½ÑÑÐ° Ð¾ÑÐ½Ð¾Ð²Ð½Ð°Ñ ÑÐµÐ»Ñ ÑÐ°Ð±Ð¾ÑÑ: Ð±ÑÐ» Ð¿Ð¾Ð»ÑÑÐµÐ½ Ð¾ÑÐ¸ÑÐµÐ½Ð½ÑÐ¹ Ð¿ÑÐµÐ¿Ð°ÑÐ°Ñ ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð½ÑÐ½Ð¾Ð³Ð¾ Ð±ÐµÐ»ÐºÐ° DprA.
description: The given work is devoted to the expression and purification of recombinant DNA-binding protein DprA from various strains of microorganisms. The tasks of the study: 1. To optimize the conditions of expression of the DprA protein gene from Escherichia coli fused with a sequence encoding 6 consecutive histidines in BL21(DE3) cells. 2. Working out of the 6his-DprA (E. c.) protein isolation scheme by chromatography methods. 3. Working out the conditions for the expression of the DprA protein gene from the Bacillus subtilis organism fused with a sequence encoding 6 consecutive genes in BL21(DE3)pLysE cells. 4. Working out of the 6his-DprA (B. s.) protein isolation scheme by chromatography methods. The object of the study is the recombinant protein DprA. The work was carried out on the basis of the laboratory of the research complex "Nanobiotechnology" as part of Peter the Great St. Petersburg Polytechnic University. Basic microbiological methods, methods of molecular biology and chromatographic protein isolation methods, as well as various methods of control and analysis were used in the work. During the research, the main goal of the work was achieved: the protein DprA was purified.