title: Ð¤ÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð°Ñ ÑÐ¸ÑÑÐµÐ¼Ð° Ð²Ð¾ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð¸Ñ ÐÐ¢Ð¤ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð°Ð´ÐµÐ½Ð¸Ð»Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Ð¸ Ð¿Ð¸ÑÑÐ²Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Escherichia coli
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: Enzymatic system of ATP regeneration based on adenylate kinase and pyruvate kinase of Escherichia coli
Creators: name: ÐÑÐ´ÑÑÐ²ÑÐµÐ²Ð°, ÐÐ°Ñ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: 2024
Subjects: ÐÐ¢Ð¤
Ð°Ð´ÐµÐ½Ð¸Ð»Ð°ÑÐºÐ¸Ð½Ð°Ð·Ð°
Ð¿Ð¸ÑÑÐ²Ð°ÑÐºÐ¸Ð½Ð°Ð·Ð°
ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð·Ð½Ð°Ñ Ð¿Ð¾Ð»Ð¸Ð¼ÐµÑÐ°Ð·Ð½Ð°Ñ Ð°Ð¼Ð¿Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ñ
ATP
adenylate kinase
pyruvate kinase
recombinase polymerase amplification

description: Ð Ð°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¿Ð¾Ð»ÑÑÐµÐ½Ð¸Ñ ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð¾Ð¹ ÑÐ¸ÑÑÐµÐ¼Ñ Ð²Ð¾ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð¸Ñ ÐÐ¢Ð¤ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð¿Ð¸ÑÑÐ²Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Ð¸ Ð°Ð´ÐµÐ½Ð¸Ð»Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Escherichia coli. Ð ÑÐ°Ð±Ð¾ÑÐµ Ð±ÑÐ»Ð¸ ÑÐºÐ¾Ð½ÑÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð½Ñ Ð¿Ð»Ð°Ð·Ð¼Ð¸Ð´Ñ, ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ¸Ðµ Ð³ÐµÐ½Ñ Ð°Ð´ÐµÐ½Ð¸Ð»Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Ð¸ Ð¿Ð¸ÑÑÐ²Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ E. coli, Ð½Ð°ÑÐ°Ð±Ð¾ÑÐ°Ð½Ñ Ð¸ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ñ ÑÐ¾Ð¾ÑÐ²ÐµÑÑÑÐ²ÑÑÑÐ¸Ðµ Ð±ÐµÐ»ÐºÐ¸. ÐÑÐ»Ð° Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð° ÐºÐ°ÑÐµÑÑÐ²ÐµÐ½Ð½Ð°Ñ Ð¿ÑÐ¾Ð²ÐµÑÐºÐ° ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð¾Ð¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð½ÑÑ Ð°Ð´ÐµÐ½Ð¸Ð»Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Ð¸ Ð¿Ð¸ÑÑÐ²Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Ð¸ ÐºÐ¾Ð»Ð¸ÑÐµÑÑÐ²ÐµÐ½Ð½Ð°Ñ Ð¿ÑÐ¾Ð²ÐµÑÐºÐ° ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð¾Ð¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð°Ð´ÐµÐ½Ð¸Ð»Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ. ÐÐ»Ñ Ð¿ÑÐ¾Ð²ÐµÑÐºÐ¸ ÑÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð¾Ð¹ ÑÐ¸ÑÑÐµÐ¼Ñ Ð²Ð¾ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð¸Ñ ÐÐ¢Ð¤ Ð±ÑÐ»Ð¾ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¾ Ð´Ð¾Ð±Ð°Ð²Ð»ÐµÐ½Ð¸Ðµ ÑÑÐ¾Ð¹ ÑÐ¸ÑÑÐµÐ¼Ñ Ðº ÑÐµÐ°ÐºÑÐ¸Ð¸ ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð·Ð½Ð¾Ð¹ Ð¿Ð¾Ð»Ð¸Ð¼ÐµÑÐ°Ð·Ð½Ð¾Ð¹ Ð°Ð¼Ð¿Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ (Ð ÐÐ), Ð¿ÑÐ¸ Ð¿ÑÐ¾ÑÐµÐºÐ°Ð½Ð¸Ð¸ ÐºÐ¾ÑÐ¾ÑÐ¾Ð¹ Ð¿Ð¾ÑÑÐµÐ±Ð»ÑÐµÑÑÑ ÐÐ¢Ð¤. ÐÐ°ÑÐµÑÑÐ²ÐµÐ½Ð½Ð°Ñ Ð¿ÑÐ¾Ð²ÐµÑÐºÐ° ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð¾Ð¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð¿Ð¸ÑÑÐ²Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Ð¿Ð¾Ð·Ð²Ð¾Ð»Ð¸Ð»Ð° ÑÐ´ÐµÐ»Ð°ÑÑ Ð²ÑÐ²Ð¾Ð´, ÑÑÐ¾ Ð¿Ð¸ÑÑÐ²Ð°ÑÐºÐ¸Ð½Ð°Ð·Ð° Ð¿ÑÐ¾ÑÐ²Ð»ÑÐµÑ ÑÐ¸Ð»ÑÐ½ÑÑ ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½ÑÑ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ. ÐÐ°ÑÐµÑÑÐ²ÐµÐ½Ð½Ð°Ñ Ð¸ ÐºÐ¾Ð»Ð¸ÑÐµÑÑÐ²ÐµÐ½Ð½Ð°Ñ Ð¿ÑÐ¾Ð²ÐµÑÐºÐ¸ ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð¾Ð¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð°Ð´ÐµÐ½Ð¸Ð»Ð°ÑÐºÐ¸Ð½Ð°Ð·Ñ Ð¿Ð¾Ð·Ð²Ð¾Ð»Ð¸Ð»Ð° ÑÐ´ÐµÐ»Ð°ÑÑ Ð²ÑÐ²Ð¾Ð´, ÑÑÐ¾ Ð°Ð´ÐµÐ½Ð¸Ð»Ð°ÑÐºÐ¸Ð½Ð°Ð·Ð° Ð¿ÑÐ¾ÑÐ²Ð»ÑÐµÑ Ð±Ð¾Ð»ÐµÐµ ÑÐ»Ð°Ð±ÑÑ, Ð½Ð¾ Ð¾Ð¿ÑÐµÐ´ÐµÐ»ÑÐµÐ¼ÑÑ ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½ÑÑ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ. Ð¡ÑÐ°Ð²Ð½ÐµÐ½Ð¸Ðµ ÑÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð°Ð¼Ð¿Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Ð² ÑÐµÐ°ÐºÑÐ¸Ð¸ Ð ÐÐ Ð±ÐµÐ· ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð¾Ð¹ ÑÐ¸ÑÑÐµÐ¼Ñ Ð²Ð¾ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð¸Ñ ÐÐ¢Ð¤ Ð¸ Ñ Ð½ÐµÐ¹, Ð¿ÑÐ¾Ð²ÐµÐ´ÑÐ½Ð½Ð¾Ðµ Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ Ð¿Ð¾Ð»Ð¸Ð¼ÐµÑÐ°Ð·Ð½Ð¾Ð¹ ÑÐµÐ¿Ð½Ð¾Ð¹ ÑÐµÐ°ÐºÑÐ¸Ð¸ Ð² ÑÐµÐ°Ð»ÑÐ½Ð¾Ð¼ Ð²ÑÐµÐ¼ÐµÐ½Ð¸, Ð¿Ð¾Ð·Ð²Ð¾Ð»Ð¸Ð»Ð¾ ÑÐ´ÐµÐ»Ð°ÑÑ Ð²ÑÐ²Ð¾Ð´, ÑÑÐ¾ ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð°Ñ ÑÐ¸ÑÑÐµÐ¼Ð° Ð²Ð¾ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð¸Ñ ÐÐ¢Ð¤ Ð¿Ð¾Ð²ÑÑÐ°ÐµÑ ÑÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ð ÐÐ.Â 
description: The given work is devoted to making enzymatic system of ATP regeneration based on adenylate kinase and pyruvate kinase of Escherichia coli. In the work plasmids, containing genes of adenylate kinase and pyruvate kinase of E. coli, were constructed, the corresponding proteins were accumulated and purified. Quality verification of enzymatic activity of purified adenylate kinase and pyruvate kinase and quantity verification of enzymatic activity of purified adenylate kinase were conducted. Addition of enzymatic system of ATP regeneration to recombinase polymerase amplification (RPA) which consumes ATP while it is happening was used to verify the efficiency of the system. Quality verification of enzymatic activity of pyruvate kinase allowed to make a conclusion that pyruvate kinase has strong enzymatic activity. Quality and quantity verification of enzymatic activity of adenylate kinase allowed to make a conclusion that adenylate kinase has comparatively weak enzymatic activity. Comparison of amplification efficiency in RPA reaction with addition of enzymatic system of ATP regeneration and without the system which was undertaken through real-time polymerase chain reaction allowed to make a conclusion that enzymatic system of ATP regeneration increases the efficiency of RPA.