title: ÐÐ½Ð°Ð»Ð¸Ð· CRISPR-Cas ÑÐ¸ÑÑÐµÐ¼ Ñ Ð¿ÑÐ¸ÑÐ¾Ð´Ð½ÑÑ ÑÑÐ°Ð¼Ð¼Ð¾Ð² Sinorhizobium meliloti
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: Analysis of CRISPR-Cas systems in natural strains of Sinorhizobium meliloti
Creators: name: ÐÐµÑÐ½Ð°Ðº, ÐÐ»Ð¸Ð·Ð°Ð²ÐµÑÐ°
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: ÐÐ¸Ð¾ÑÐµÑ Ð½Ð¾Ð»Ð¾Ð³Ð¸Ñ
ÑÐ°Ð³Ð¾ÑÑÑÐ¾Ð¹ÑÐ¸Ð²Ð¾ÑÑÑ
ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ Ð¿Ð¾ Ð¡ÑÐ½Ð³ÐµÑÑ
Ð±Ð¸Ð¾Ð¸Ð½ÑÐ¾ÑÐ¼Ð°ÑÐ¸ÑÐµÑÐºÐ¸Ð¹ Ð°Ð½Ð°Ð»Ð¸Ð·
phage resistance
Sanger sequencing
bioinformatics analysis

description: Ð¢ÐµÐ¼Ð° Ð²ÑÐ¿ÑÑÐºÐ½Ð¾Ð¹ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÑ: Â«ÐÐ½Ð°Ð»Ð¸Ð· CRISPR-Cas ÑÐ¸ÑÑÐµÐ¼ Ñ Ð¿ÑÐ¸ÑÐ¾Ð´Ð½ÑÑ ÑÑÐ°Ð¼Ð¼Ð¾Ð² Sinorhizobium melilotiÂ». ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ ÑÐ°Ð·Ð½Ð¾Ð¾Ð±ÑÐ°Ð·Ð¸Ñ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½ÑÑ ÑÐ»ÐµÐ¼ÐµÐ½ÑÐ¾Ð² CRISPR-Cas ÑÐ¸ÑÑÐµÐ¼ Ð² Ð¿ÑÐ¸ÑÐ¾Ð´Ð½ÑÑ Ð¿Ð¾Ð¿ÑÐ»ÑÑÐ¸ÑÑ S. meliloti Ð¸ Ð¾ÑÐµÐ½ÐºÐ¸ Ð¸Ñ Ð²Ð·Ð°Ð¸Ð¼Ð¾ÑÐ²ÑÐ·Ð¸ Ñ ÑÐ°Ð³Ð¾ÑÑÑÐ¾Ð¹ÑÐ¸Ð²Ð¾ÑÑÑÑ. Ð Ð°Ð±Ð¾ÑÐ° Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð° Ð½Ð° Ð±Ð°Ð·Ðµ Ð»Ð°Ð±Ð¾ÑÐ°ÑÐ¾ÑÐ¸Ð¸ Ð³ÐµÐ½ÐµÑÐ¸ÐºÐ¸ Ð¸ ÑÐµÐ»ÐµÐºÑÐ¸Ð¸ Ð¼Ð¸ÐºÑÐ¾Ð¾ÑÐ³Ð°Ð½Ð¸Ð·Ð¼Ð¾Ð² Ð¤ÐÐÐÐ£ ÐÐÐÐÐ¡Ð¥Ð, Ð³Ð´Ðµ Ð²Ð¿ÐµÑÐ²ÑÐµ Ð±ÑÐ»Ð¸ Ð¸Ð·ÑÑÐµÐ½Ñ Ð½Ð°Ð»Ð¸ÑÐ¸Ðµ, ÑÑÑÑÐºÑÑÑÐ° Ð¸ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½Ð°Ñ ÑÐ¾Ð»Ñ 7 ÑÐ»ÐµÐ¼ÐµÐ½ÑÐ¾Ð² CRISPR-Cas ÑÐ¸ÑÑÐµÐ¼ Ð² Ð³ÐµÐ½Ð¾Ð¼Ð°Ñ Ð¿ÑÐ¸ÑÐ¾Ð´Ð½ÑÑ ÑÑÐ°Ð¼Ð¼Ð¾Ð² ÑÐ¾Ð·ÑÐ¹ÑÑÐ²ÐµÐ½Ð½Ð¾-ÑÐµÐ½Ð½ÑÑ Ð°Ð·Ð¾ÑÑÐ¸ÐºÑÐ¸ÑÑÑÑÐ¸Ñ Ð±Ð°ÐºÑÐµÑÐ¸Ð¹ Ð²Ð¸Ð´Ð° S. meliloti Ð¸Ð· ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÑ ÑÐµÐ³Ð¸Ð¾Ð½Ð¾Ð² (ÐÐ°Ð²ÐºÐ°Ð·ÑÐºÐ¾Ð³Ð¾, ÐÑÐ°Ð»ÑÑÐºÐ¾Ð³Ð¾, Ð¡ÑÐµÐ´Ð½ÐµÐ°Ð·Ð¸Ð°ÑÑÐºÐ¾Ð³Ð¾ Ð¸ Ð¢ÑÑÐºÐ¼ÐµÐ½ÑÐºÐ¾Ð³Ð¾) Ñ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸ÐµÐ¼ Ð¿Ð°Ñ Ð¿ÑÐ°Ð¹Ð¼ÐµÑÐ¾Ð², ÑÐºÐ¾Ð½ÑÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ Ð² ÑÐ¾Ð´Ðµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ. ÐÐµÑÐ¾Ð´Ð¾Ð¼ ÐÐ¦Ð  Ð² 146 ÑÑÐ°Ð¼Ð¼Ð°Ñ, ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ¸Ñ 1022 Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½ÑÐµ CRISPR ÐºÐ°ÑÑÐµÑÑ, Ð±ÑÐ»Ð¾ Ð²ÑÑÐ²Ð»ÐµÐ½Ð¾ 140 (95,5%) ÑÑÐ°Ð¼Ð¼Ð¾Ð² Ñ 241 (23,6%) Ð¸Ð·Ð¼ÐµÐ½ÐµÐ½Ð½Ð¾Ð¹ CRISPR ÐºÐ°ÑÑÐµÑÐ¾Ð¹. ÐÑÐµÐ½ÐºÐ° ÑÑÑÑÐºÑÑÑÐ½Ð¾Ð³Ð¾ Ð¿Ð¾Ð»Ð¸Ð¼Ð¾ÑÑÐ¸Ð·Ð¼Ð° Ð¿Ð¾ÐºÐ°Ð·Ð°Ð»Ð°, ÑÑÐ¾ Ð¸Ð·Ð¼ÐµÐ½ÐµÐ½Ð½ÑÐµ ÐºÐ°ÑÑÐµÑÑ ÑÐ°ÑÐµ Ð²ÑÑÑÐµÑÐ°Ð»Ð¸ÑÑ Ñ Ð¿Ð¾ÑÐ²ÐµÐ½Ð½ÑÑ ÑÑÐ±Ð¿Ð¾Ð¿ÑÐ»ÑÑÐ¸Ð¹, ÑÐµÐ¼ Ñ ÐºÐ»ÑÐ±ÐµÐ½ÑÐºÐ¾Ð²ÑÑ. ÐÐ°ÑÑÐµÑÐ° Ð¡R-1, Ð±ÑÐ»Ð° Ð¸Ð·Ð¼ÐµÐ½ÐµÐ½Ð° Ñ 94,5% ÑÑÐ°Ð¼Ð¼Ð¾Ð² Ð² ÐºÐ°Ð¶Ð´Ð¾Ð¹ Ð¸Ð· Ð¿Ð¾Ð¿ÑÐ»ÑÑÐ¸Ð¹. ÐÐ°ÑÑÐµÑÑ CR-4 Ð¸ CR-5 Ð¼ÐµÐ½ÑÐ»Ð¸ÑÑ ÑÐµÐ¶Ðµ Ð¾ÑÑÐ°Ð»ÑÐ½ÑÑ, Ð»Ð¸ÑÑ Ñ 3% ÑÑÐ°Ð¼Ð¼Ð¾Ð². ÐÐ°Ð¸Ð±Ð¾Ð»ÑÑÐµÐµ ÑÐ°Ð·Ð½Ð¾Ð¾Ð±ÑÐ°Ð·Ð¸Ðµ ÑÐ¸Ð¿Ð¾Ð² ÐºÐ°ÑÑÐµÑ Ð±ÑÐ»Ð¾ Ð²ÑÑÐ²Ð»ÐµÐ½Ð¾ Ñ CR-3 (3-5 ÑÐ¸Ð¿Ð¾Ð²), Ð½Ð°Ð¸Ð¼ÐµÐ½ÑÑÐµÐµ â Ñ CR-1 (1-2 ÑÐ¸Ð¿Ð°). Ð¢Ð°ÐºÐ¶Ðµ Ð±ÑÐ»Ð° Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð° Ð¾ÑÐµÐ½ÐºÐ° ÑÐ°Ð³Ð¾ÑÑÑÐ¾Ð¹ÑÐ¸Ð²Ð¾ÑÑÐ¸ ÑÑÐ°Ð¼Ð¼Ð¾Ð², Ð¸Ð¼ÐµÑÑÐ¸Ñ Ð¸Ð·Ð¼ÐµÐ½ÐµÐ½Ð½ÑÐµ CRISPR ÐºÐ°ÑÑÐµÑÑ. ÐÑÐ»Ð¾ ÑÐ´ÐµÐ»Ð°Ð½Ð¾ Ð¿ÑÐµÐ´Ð¿Ð¾Ð»Ð¾Ð¶ÐµÐ½Ð¸Ðµ, ÑÑÐ¾ Ð½Ð°Ð¸Ð±Ð¾Ð»ÑÑÑÑ ÑÐ°Ð³Ð¾ÑÑÑÐ¾Ð¹ÑÐ¸Ð²Ð¾ÑÑÑ Ð¾Ð±ÐµÑÐ¿ÐµÑÐ¸Ð²Ð°ÐµÑ ÐºÐ°ÑÑÐµÑÐ° CR-5, Ð° Ð½Ð°Ð¸Ð¼ÐµÐ½ÑÑÑÑ â CR-6. Ð ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð° ÑÐ¾Ð·Ð´Ð°Ð½Ð° Ð±Ð°Ð·Ð° Ð´Ð°Ð½Ð½ÑÑ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾ÑÑÐµÐ¹ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½ÑÑ ÑÐ»ÐµÐ¼ÐµÐ½ÑÐ¾Ð² CRISPR-Cas ÑÐ¸ÑÑÐµÐ¼ ÑÐ°ÑÐ°ÐºÑÐµÑÐ½ÑÑ Ð´Ð»Ñ S. meliloti Ð² ÑÐ²ÑÐ·Ð¸ Ñ ÑÐµÐ·Ð¸ÑÑÐµÐ½ÑÐ½Ð¾ÑÑÑÑ Ðº ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÐ¼ Ð±Ð°ÐºÑÐµÑÐ¸Ð¾ÑÐ°Ð³Ð°Ð¼.
description: Topic of the final qualification work: "Analysis of CRISPR-Cas systems in natural strains of Sinorhizobium meliloti". This work is devoted to the study of the diversity of potential elements of CRISPR-Cas systems in natural populations of S. meliloti and the assessment of their relationship with phage resistance. The work was carried out on the basis of the Laboratory of Genetics and Breeding of Microorganisms of the FSBSI ARRIAM, where studied for the first time the presence, structure and potential role of 7 elements of CRISPR-Cas systems in the genomes of natural strains of economically valuable nitrogen-fixing bacteria of the species S. meliloti from various regions (Caucasian, Aral, Central Asian and Turkmensky) with using pairs of primers designed during the study. By PCR in 146 strains containing 1022 potential CRISPR cassettes revealed 140 (95.5%) strains with 241 (23.6%) altered CRISPR cassettes. An assessment of structural polymorphism showed that altered cassettes were more common in soil subpopulations than in nodule subpopulations. The CR-1 cassette was altered in 94.5% of the strains in each population. The CR-4 and CR-5 cassettes changed less frequently than others, only in 3% of the strains. The greatest diversity of cassette types was found in CR-3 (types 3-5), the smallest in CR-1 (types 1-2). The phage resistance of strains with altered CRISPR cassettes was also assessed. It was assumed that the CR-5 cassette provides the greatest phage resistance, and the CR-6 cassette provides the least. As a result of the work, a database of sequences of potential elements of CRISPR-Cas systems characteristic of S. meliloti in connection with resistance to various bacteriophages was created.