Genetic knowledge of microorganisms plays a critical role in the creation of new biotechnologies, since the effectiveness of any biotechnology is determined by the particular qualities of the structurally functional organization of molecular-genetic systems and their components used for the production of targeted products. Collections of microbial cultures play a decisive role in mobilizing biological resources and make it possible to form a solid base for genetic, molecular biological and biotechnological research. The aim of this work was to assess the key molecular-genetic and phenotypic characteristics of strains of the collection of microorganisms created in the "FRC Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences" as a genetic resource for biotechnology. Thirty strains of microorganisms of the collection were isolated by employees of the FRC ICG SB RAS from extreme natural ecosystems, the key molecular-genetic and phenotypic characteristics were described using modern methods of molecular biology and mass-spectrometry. DNA isolation and the sequencing of 16S rRNA gene sequences were performed. The strains of the collection were characterized by morphological, physiological, moleculargenetic and mass-spectrometric characteristics. The particular qualities of growing of strains on different substrates have been established, the study of cell morphology has been carried out. The physiological characteristics of the strains of the collection have been established: the attitude to oxygen, the type of nutrition, the range of temperature and pH, the attitude to NaCl and others. Different resistance of strains to antibiotics has been established. The creation of personal mass spectra of protein profiles of the studied strains of the collection was carried out. The resulting DNA sequences of the strains are deposited in the GenBank. The chemotaxonomic characteristics В создании новых биотехнологий важное значение имеют генети-ческие знания о микроорганизмах, так как эффективность любых биотехнологий определяется особенностями структурно-функ-циональной организации молекулярно-генетических систем и их компонент, используемых для наработки целевых продуктов. При этом коллекции микробных культур играют решающую роль в мо-билизации биологических ресурсов и позволяют сформировать прочную базу для генетических, молекулярно-биологических и биотехнологических исследований. Целью данной работы было проведение оценки ключевых молекулярно-генетических и фено-типических характеристик штаммов коллекции микроорганизмов, созданной в Федеральном исследовательском центре Институт цитологии и генетики (ФИЦ ИЦиГ) СО РАН, в качестве генетическо-го ресурса для биотехнологии. Для 30 штаммов микроорганизмов коллекции, выделенных сотрудниками ИЦиГ из экстремальных природных экосистем, осуществлено описание ключевых молеку-лярно-генетических и фенотипических характеристик с использо-ванием современных методов молекулярной биологии и масс-спектрометрии. Проведено выделение ДНК и секвенирование по...
Yeasts are a convenient model eukaryote used for genome studies and genome editing. Saccharomyces cerevisiae is the species most widely employed in bio technology, since it is easily cultivated in bioreactors and is absolutely safe. The last decade saw a significant development of methods of yeast genetic engineer ing and the creation of novel instruments adapted from other fields, which allowed one to significantly accelerate the construction of new strains. The most prominent examples are the proteins used for directed DNA editing. For a long time, yeast genome engineer ing was based on the yeasts' system of homologous recombination. It was sufficient for several decades before the development of highthroughput methods. Many highthroughput methods were developed in the second decade of the XXI century, including those used in genomics, transcriptomics, proteomics, metabolomics, interactomics, etc. Modern bioinfor matic databases now allow one to rapidly process the increasing flow of information and model cellular pro cesses. As a result, the rate of analysis and prediction of targets for genome editing is currently higher than the rate of genome editing, which led to the develop ment of new methods of genetic engineering. This process was particularly pronounced for microorgan isms. Modern tasks require tens, hundreds, sometimes even thousands of genome modifications, which made researchers to look for new techniques. As a result, the instruments used for more complex objects, such as animals, plants, and cell lines, were adapted for yeasts. Modern methods for yeast genome editing allow introducing several modifications into the genome in a single step. In this study, we review the methods of directed genome editing and their applications and perspectives for yeasts.Key words: yeast; Saccharomyces cerevisiae; genetic engineering; ZFN; zinc fingers; TALENS; CRISPR/Cas; Argonaut; NgAgo.Дрожжи являются модельным эукариотическим организмом, на котором отрабатываются многие предположения о работе генома, а также методы его редактирования. Наиболее часто в исследо вательских работах используют Saccharomyces cerevisiae, которые очень хорошо приспособлены физиологически к культивированию в условиях биореактора и признаны абсолютно безопасными. В по следнее десятилетие методы генетической инженерии дрожжей претерпели значительные изменения. Появились новые инстру менты, которые пришли из смежных направлений и позволили значительно ускорить процесс получения новых штаммов. Прежде всего это белки для направленного внесения изменений в после довательность ДНК. Длительное время методы редактирования генома дрожжей базировались на использовании их собственной системы гомологичной рекомбинации. Она удобна и удовлетво ряла потребности исследователей на протяжении нескольких де сятилетий, до того времени, когда на первый план стали выходить высокопроизводительные методы. Во втором десятилетии XXI века произошло бурное развитие высокопроизводительных подходов, в первую очередь методов анализа в биологии: геномики, транс крипто...
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