Efficient genome editing of an extreme thermophile, Thermus thermophilus, using a thermostable Cas9 variant
نویسندگان
چکیده
منابع مشابه
A plasmid vector for an extreme thermophile, Thermus thermophilus.
The host-vector system for an extreme thermophile, Thermus thermophilus HB27, was developed. The host strain has a mutation in tryptophan synthetase gene (trpB), and the mutation was determined to be a missense mutation by DNA sequence analysis. A Thermus-E. coli shuttle vector pYK109 was constructed. pYK109 consists of Thermus cryptic plasmid pTT8, tryptophan synthetase gene (trpB) of Thermus ...
متن کاملAn extreme thermophile, Thermus thermophilus, is a polyploid bacterium.
An extremely thermophilic bacterium, Thermus thermophilus HB8, is one of the model organisms for systems biology. Its genome consists of a chromosome (1.85 Mb), a megaplasmid (0.26 Mb) designated pTT27, and a plasmid (9.3 kb) designated pTT8, and the complete sequence is available. We show here that T. thermophilus is a polyploid organism, harboring multiple genomic copies in a cell. In the cas...
متن کاملFunctions of isolated domains of methionyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8.
Methionyl-tRNA synthetase (MetRS, 2 X 75 kDa) was purified to homogeneity from an extreme thermophile, Thermus thermophilus HB8. The polypeptide chain of MetRS was cleaved by limited digestion with trypsin into four domains: T1 (29 kDa), T2 (23 kDa), T3 (14.5 kDa), and T4 (7.5 kDa), which were aligned in that order. MetRS was also cleaved into similar fragments with a variety of other proteases...
متن کاملEfficient Genome Editing of a Facultative Thermophile Using Mesophilic spCas9
Well-developed genetic tools for thermophilic microorganisms are scarce, despite their industrial and scientific relevance. Whereas highly efficient CRISPR/Cas9-based genome editing is on the rise in prokaryotes, it has never been employed in a thermophile. Here, we apply Streptococcus pyogenes Cas9 (spCas9)-based genome editing to a moderate thermophile, i.e., Bacillus smithii, including a gen...
متن کاملThermally induced biosynthesis of 2'-O-methylguanosine in tRNA from an extreme thermophile, Thermus thermophilus HB27.
The contents of 2'-O-methylguanosine and 1-methyladenosine in unfractionated tRNA obtained from Thermus thermophilus HB27 were found to increase significantly when the bacterium was grown at a higher temperature (80 degrees C). S-Adenosyl-L-methionine-dependent tRNA (guanosine-2')-methyltransferase (EC 2.1.1.34) and tRNA (adenine-1)-methyltransferase (EC 2.1.1.36) were detected in a cell-free e...
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ژورنال
عنوان ژورنال: Scientific Reports
سال: 2021
ISSN: 2045-2322
DOI: 10.1038/s41598-021-89029-2