A bank of Streptomyces plasmid pSS27 fragments was created on the basis of the vector pBluescript II SK(+) and a bifunctional vector pWHM4. Strains of Escherichia coli DH1 and JM109 were used as recipients of hybrid DNA. Selected transformants contained hybrid molecules of DNA with inserts of Streptomyces plasmid from 0.5 kb up to 12.3 kb. Some thiostrepton-resistant transformants which contained extrachromosomal DNA with the molecular size of 18.1 kb were obtained by means of transformation by hybrid plasmid DNA pSW51 (18.1 kb) of S. levoris 165 protoplasts. The analysis of nucleotide sequences of some hybrid plasmids has shown homology of sequencing parts of cloned fragments of plasmid pSS27 with sequences of fibers (chromosomal genes), which took part in sporulation of S. coelicolor A3 (2) and S. avermitilis MA-4680 and Tra B gene of S. phaeochromogenes plasmid pJV1.
Download full-text PDF |
Source |
---|
Sci China Life Sci
January 2025
Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China.
Since their discovery, CRISPR/Cas systems have significantly expanded the genetic toolbox, aiding in the exploration and enhanced production of natural products across various microbes. Among these, class 2 CRISPR/Cas systems are simpler and more broadly used, but they frequently fail to function effectively in many Streptomyces strains. In this study, we present an engineered class 1 type I CRISPR/Cas system derived from Streptomyces avermitilis, which enables efficient gene editing in phylogenetically distant Streptomyces strains.
View Article and Find Full Text PDFMicrobiol Resour Announc
January 2025
School of Natural and Environmental Sciences, Newcastle University, Newcastle upon Tyne, United Kingdom.
Here, we report the resequencing, assembly, and annotation of two actinomycete genomes containing abyssomicin gene clusters. DSM 45791 with a circular chromosome of 11,681,598 bp and 4 circular plasmids (14,175-207,548 bp) and sp. NL15-2K with a 12,368,159 bp linear genome and circular plasmid (11,584 bp).
View Article and Find Full Text PDFAntibiotics (Basel)
November 2024
Department of Chemical Engineering and Environmental Technology, School of Industrial Engineering, University of Valladolid, Paseo Prado de la Magdalena 3-5, 47011 Valladolid, Spain.
CRISPR/Cas systems have emerged as valuable tools to approach the problem of antimicrobial resistance by either sensitizing or lysing resistant bacteria or by aiding in antibiotic development, with successful applications across diverse organisms, including bacteria and fungi. CRISPR/Cas systems can target plasmids or the bacterial chromosome of AMR-bacteria, and it is especially necessary to have an efficient entry into the target cells, which can be achieved through nanoparticles or bacteriophages. Regarding antibiotic development and production, though the use of CRISPR/Cas in this field is still modest, there is an untapped reservoir of bacterial and fungal natural products, with over 95% yet to be characterized.
View Article and Find Full Text PDFAppl Environ Microbiol
December 2024
MOE Key Laboratory of Industrial Fermentation Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, China.
Over the past three decades, the integrase (Int) from phage C31 has become a valuable genome engineering tool across various species. C31 Int was thought to mediate unidirectional site-specific integration ( × to and ) in the absence of the phage-encoded recombination directionality factor (RDF). However, we have shown in this study that Int can also catalyze reverse excision ( × to and ) at low frequencies in and , causing genetic instability in engineered strains.
View Article and Find Full Text PDFCurr Microbiol
December 2024
Faculty of Science, Molecular Biology and Genetics Department, Gebze Technical University, Gebze, 41400, Kocaeli, Türkiye.
The genus Streptomyces is a group of gram-positive bacteria that exhibit a distinctive growth pattern characterised by elongated, branched hyphae. Streptomyces coelicolor A3(2), which produces at least five different antibiotics, is a model organism that is widely used in genetic studies. There are very few studies in Streptomyces on the ATP-dependent Lon protease, which has very important functions in every organism and is particularly responsible for protein homeostasis.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!