title: ÐÐ·ÑÑÐµÐ½Ð¸Ðµ Ð³Ð¸Ð³Ð°Ð½ÑÑÐºÐ¸Ñ Cas1-Ð¿Ð¾Ð´Ð¾Ð±Ð½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð² Ð¸Ð· Asgard Ð°ÑÑ ÐµÐ¹
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: Studying of giant Cas1-like proteins from Asgard Ð°rchaea
Creators: name: ÐÐ°Ð»ÑÑ, ÐÐ½Ð°ÑÑ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: 2023
Subjects: cas1
Ð°ÑÑ ÐµÐ¸
ÑÑÐ°Ð½ÑÐ¿Ð¾Ð·Ð¾Ð½Ñ
Ð¼Ð¾Ð±Ð¸Ð»ÑÐ½ÑÐµ Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¸Ðµ ÑÐ»ÐµÐ¼ÐµÐ½ÑÑ
archaea
transposons
mobile genetic elements

description: ÐÐ°Ð´Ð°ÑÐ¸ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ: - ÑÐ¾Ð·Ð´Ð°ÑÑ Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÑÑ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ñ, ÐºÐ¾Ð´Ð¸ÑÑÑÑÑÑ Ð³ÐµÐ½ Ð±ÐµÐ»ÐºÐ° Cas1_3 Ñ ÑÐ°Ð³Ð¾Ð¼ MBP. - Ð¿Ð¾Ð»ÑÑÐ¸ÑÑ ÑÐ¸ÑÑÑÐ¹ ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð½ÑÐ½ÑÐ¹ Ð±ÐµÐ»Ð¾Ðº Cas1_3 Ð¼ÐµÑÐ¾Ð´Ð°Ð¼Ð¸ Ð°ÑÑÐ¸Ð½Ð½Ð¾Ð¹ Ð¸ Ð³ÐµÐ»Ñ-ÑÐ¸Ð»ÑÑÑÐ°ÑÐ¸Ð¾Ð½Ð½Ð¾Ð¹ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ð¸. - Ð¾Ð¿ÑÐµÐ´ÐµÐ»Ð¸ÑÑ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½ÑÐµ Ð´Ð¾Ð¼ÐµÐ½Ñ Ð±ÐµÐ»ÐºÐ° Cas1_3 Ð¿ÑÑÐµÐ¼ Ð¿Ð¾Ð¸ÑÐºÐ° Ð³Ð¾Ð¼Ð¾Ð»Ð¾-Ð³Ð¸ÑÐ½ÑÑ ÐµÐ¼Ñ Ð´Ð¾Ð¼ÐµÐ½Ð¾Ð² Ð´ÑÑÐ³Ð¸Ñ Ð±ÐµÐ»ÐºÐ¾Ð², ÑÑÐ½ÐºÑÐ¸Ñ ÐºÐ¾ÑÐ¾ÑÑÑ Ð¸Ð·Ð²ÐµÑÑÐ½Ð°, Ð¸ÑÐ¿Ð¾Ð»ÑÐ·ÑÑ ÐºÐ¾Ð¼-Ð¿ÑÑÑÐµÑÐ½Ð¾Ðµ Ð¼Ð¾Ð´ÐµÐ»Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ ÑÑÑÑÐºÑÑÑÑ. - Ð² Ð·Ð°Ð²Ð¸ÑÐ¸Ð¼Ð¾ÑÑÐ¸ Ð¾Ñ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾ÑÑÐ¸ Ð½Ð°Ð¹Ð´ÐµÐ½Ð½ÑÑ Ð² Cas1_3 Ð±ÐµÐ»ÐºÐµ Ð´Ð¾Ð¼ÐµÐ½Ð¾Ð² Ð¿ÑÐ¾Ð²ÐµÑÐ¸ÑÑ Ð¸Ñ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ in vitro ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½ÑÐ¾Ð². ÐÑÐµÐ´Ð¼ÐµÑÐ¾Ð¼ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ ÑÐ²Ð»ÑÑÑÑÑ ÑÑÑÑÐºÑÑÑÐ° Ð¸ ÑÐ²Ð¾Ð¹ÑÑÐ²Ð° Ð³Ð¸Ð³Ð°Ð½ÑÑÐºÐ¾Ð³Ð¾ Cas1-Ð¿Ð¾Ð´Ð¾Ð±Ð½Ð¾Ð³Ð¾ Ð±ÐµÐ»ÐºÐ°, ÑÐ²Ð»ÑÑÑÐµÐ³Ð¾ÑÑ Ð¾ÑÐ½Ð¾Ð²Ð½ÑÐ¼ ÐºÐ¾Ð¼Ð¿Ð¾Ð½ÐµÐ½ÑÐ¾Ð¼ Ð¿ÑÐµÐ´Ð¿Ð¾Ð»Ð¾Ð¶Ð¸ÑÐµÐ»ÑÐ½Ð¾ ÑÑÐ°Ð½ÑÐ¿Ð¾Ð·Ð°Ð±ÐµÐ»ÑÐ½ÑÑ Ð¸ Ð·Ð°ÑÐ¸ÑÐ½ÑÑ Cas1-ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ¸Ñ ÑÐ¸ÑÑÐµÐ¼ (Ð´Ð°Ð»ÐµÐµ â Cas1_3 ÑÐ¸ÑÑÐµÐ¼Ñ) Ð¸Ð· Asgard Ð°ÑÑÐµÐ¹. Ð Ð°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¸Ð·ÑÑÐµÐ½Ð¸Ñ Ð±ÐµÐ»ÐºÐ° Cas1_3, Ð³ÐµÐ½ ÐºÐ¾ÑÐ¾ÑÐ¾Ð³Ð¾ Ð±ÑÐ» Ð¾Ð±Ð½Ð°ÑÑÐ¶ÐµÐ½ Ð² Ð¼ÐµÑÐ°Ð³ÐµÐ½Ð¾Ð¼Ðµ Asgard Ð°ÑÑÐµÐ¹ Ð² ÑÐ¾ÑÑÐ°Ð²Ðµ ÑÐ¸ÑÑÐµÐ¼Ñ-ÑÐ²Ð¾Ð»ÑÑÐ¸Ð¾Ð½Ð½Ð¾Ð³Ð¾ Ð¿ÑÐµÐ´ÑÐµÑÑÐ²ÐµÐ½Ð½Ð¸ÐºÐ° CRISPR-Cas ÑÐ¸ÑÑÐµÐ¼ II ÐºÐ»Ð°ÑÑÐ°. ÐÐ±Ð·Ð¾Ñ Ð»Ð¸ÑÐµÑÐ°ÑÑÑÑ Ð²ÐºÐ»ÑÑÐ°ÐµÑ Ð¾Ð¿Ð¸ÑÐ°Ð½Ð¸Ðµ Ð·Ð°ÑÐ¸ÑÐ½ÑÑ ÑÐ¸ÑÑÐµÐ¼ Ð¿ÑÐ¾ÐºÐ°ÑÐ¸Ð¾Ñ, Ð¸Ñ Ð¾ÑÐ³Ð°Ð½Ð¸Ð·Ð°ÑÐ¸Ð¸ Ð² Ð·Ð°ÑÐ¸ÑÐ½ÑÐµ Ð¾ÑÑÑÐ¾Ð²ÐºÐ¸, Ð° ÑÐ°ÐºÐ¶Ðµ ÑÐ°ÑÐ°ÐºÑÐµÑÐ¸ÑÑÐ¸ÐºÐ° Ð¸Ð·ÑÑÐ°ÐµÐ¼Ð¾Ð¹ Ð°ÑÑÐµÐ¹Ð½Ð¾Ð¹ ÑÐ¸ÑÑÐµÐ¼Ñ Ð¸ ÐµÐµ ÐºÐ¾Ð¼Ð¿Ð¾Ð½ÐµÐ½ÑÐ¾Ð². ÐÑÐ¸ Ð½Ð°Ð¿Ð¸ÑÐ°Ð½Ð¸Ð¸ Ð»Ð¸ÑÐµÑÐ°ÑÑÑÐ½Ð¾Ð³Ð¾ Ð¾Ð±Ð·Ð¾ÑÐ° Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð»Ð¸ÑÑ Ð±Ð°Ð·Ñ Ð´Ð°Ð½Ð½ÑÑ NCBI, Scopus Ð¸ ScienceDirect. ÐÑÐ»Ð° ÑÐ¼Ð¾Ð´ÐµÐ»Ð¸ÑÐ¾Ð²Ð°Ð½Ð° ÑÑÐµÑÐ¼ÐµÑÐ½Ð°Ñ ÑÑÑÑÐºÑÑÑÐ° Ð±ÐµÐ»ÐºÐ° Cas1_3; Ð¾Ð¿ÑÐµÐ´ÐµÐ»ÐµÐ½Ñ ÐµÐ³Ð¾ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½ÑÐµ Ð´Ð¾Ð¼ÐµÐ½Ñ Ð¸ Ð¿ÑÐµÐ´Ð¿Ð¾Ð»Ð¾Ð¶ÐµÐ½Ñ Ð±Ð¸Ð¾Ð»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¸Ðµ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸. ÐÑÐ´ÐµÐ»ÐµÐ½ ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð½ÑÐ½ÑÐ¹ Ð±ÐµÐ»Ð¾Ðº Cas1_3, ÑÐ»Ð¸ÑÑÐ¹ Ñ MBP, Ð¸ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ñ in vitro ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½ÑÑ Ð¿Ð¾ Ð¿ÑÐ¾Ð²ÐµÑÐºÐµ Ð½Ð°Ð»Ð¸ÑÐ¸Ñ Ñ Ð½ÐµÐ³Ð¾ ÐÐ¢Ð¤Ð°Ð·Ð½Ð¾Ð¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð¸ Ð²Ð¾Ð·Ð¼Ð¾Ð¶Ð½Ð¾ÑÑÐ¸ ÑÐ²ÑÐ·ÑÐ²Ð°Ð½Ð¸Ñ Ñ Ð½ÑÐºÐ»ÐµÐ¸Ð½Ð¾Ð²ÑÐ¼Ð¸ ÐºÐ¸ÑÐ»Ð¾ÑÐ°Ð¼Ð¸. Ð¡ÑÐ°ÑÐ¸ÑÑÐ¸ÑÐµÑÐºÑÑ Ð¾Ð±ÑÐ°Ð±Ð¾ÑÐºÑ ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½ÑÐ°Ð»ÑÐ½ÑÑ Ð´Ð°Ð½Ð½ÑÑ Ð¿ÑÐ¾Ð²Ð¾Ð´Ð¸Ð»Ð¸ Ñ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸ÐµÐ¼ Ð¿Ð°ÐºÐµÑÐ° Ð¿ÑÐ¾Ð³ÑÐ°Ð¼Ð¼ Microsoft Excel, Origin.
description: Study objectives: - to create a genetic construction encoding the Cas1_3 protein gene with MBP tag. - to purify recombinant Cas1_3 protein using affinity and size-exclusion chro-matography. - to determine Cas1_3 functional domains using computer structural modeling. - based on the proposed functions of the domains predicted in Cas1_3 protein, test the corresponding activities in vitro. The subject of the study is structure and functions of a giant Cas1-like protein, the main component of transposable and presumably defensive Cas1-containing systems from Asgard archaea (further â Cas1_3 systems). This work is focused on the studying of Cas1_3 protein, found in the Asgard archaea metagenome as a part of the system â the evolutionary precursor of CRISPR-Cas class II systems. The description of the defense systems of prokaryotes, their organization into defense islands, as well as the dicription of the archaeal Cas1-containing putative transposable systems and its components, the giant Cas1-like protein and the TnpB protein, are presented. NCBI, Scopus, and ScienceDirect databases were used for the subject overview. The three-dimensional structure of Cas1_3 protein was modeled partially; the functional domains and putative functions of the protein were determined. The recombinant Cas1_3_MBP protein was purified, and in vitro experiments were carried out to detect ATPase activity and proteinâs binding with nucleic acids. Statistics were processed using Microsoft Excel, Origin software package.