17 results about "Nucleotide substitution" patented technology

The invention relates to plant lactobacillus and application thereof. Wherein the gene of the molecular marker for the plant lactobacillus has a nucleotide sequence selected from one of the following sequences: (1) a nucleotide sequence as shown in SEQ ID NO: 24; (2) a nucleotide sequence having at least 97%, 98%, 99% or higher homology with the nucleotide sequence as shown in SEQ ID NO: 24; and (3) a nucleotide sequence with one or more nucleotide sequences, such as 1, 2, 3, 4, 5 or more nucleotide substitutions, deletion or insertion, in the nucleotide sequence as shown in SEQ ID NO: 24. The plant lactobacillus provided by the invention is non-toxic, has good biological characteristics, has a good curative effect on prevention and / or treatment of vaginal infection and related diseases, and avoids the problems of increased drug resistance, high recurrence rate and the like caused by use of antibiotic and other antibacterial drugs during treatment of vaginal infection and related diseases.

In traditional plant breeding approaches, chemical mutagenesis may be utilized to introduce nucleotide substitutions at random in the genome of a plant, i.e. without possibilities to control the sites of nucleotide changes. Because of genome complexities, the statistical probability is extremely little when it comes to finding a predetermined nucleotide substitution. The present invention, however, demonstrates how a novel, alternative use of digital polymerase chain reaction (dPCR), preferably droplet dPCR (ddPCR), is developed to exploit finding of specific nucleotide substitutions in mutated genes. The entire platform comprises a screening method with a library of mutagenized organisms, digital PCR-based systems and a set-up to propagate and analyze identified, mutated organisms.

The present invention provides Lactiplantibacillus plantarum and a use thereof. A molecular marker gene for Lactiplantibacillus plantarum has a nucleotide sequence selected from one of the following: (1) a nucleotide sequence as shown in SEQ ID NO: 24; (2) a nucleotide sequence having at least 97%, 98%, 99% or higher homology with the nucleotide sequence as shown in SEQ ID NO: 24; and (3) a nucleotide sequence having one or more, e.g., 1, 2, 3, 4, 5, or more, nucleotide substitutions, deletions, or insertions in the nucleotide sequence as shown in SEQ ID NO: 24. The Lactiplantibacillus plantarum provided by the present invention has non-toxicity and good biological characteristics and has a good curative effect on the prevention and / or treatment of vaginal infection and related diseases thereof.

This invention proposes a molecular marker for *Lactobacillus japonicus*, which was deposited on February 7, 2023, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 26505. The molecular marker comprises a nucleotide sequence selected from one of the following: (1) the nucleotide sequence shown in SEQ ID NO: 24; (2) a nucleotide sequence having at least 97%, 98%, 99%, or higher homology to the nucleotide sequence shown in SEQ ID NO: 24; or (3) a nucleotide sequence having one or more nucleotide substitutions, deletions, or insertions in the nucleotide sequence shown in SEQ ID NO: 24. Using the molecular marker of this invention, not only can the target *Lactobacillus japonicus* be screened quickly and easily, but it can also highly specifically identify the target *Lactobacillus japonicus*, eliminating interference from other strains.

The invention relates to application of clostridium butyricum in preparation of a product for improving sleep, and a gene sequence of the clostridium butyricum comprises one of the following nucleotide sequences: (1) a nucleotide sequence as shown in SEQ ID NO: 1; (2) a nucleotide sequence having at least 99.8%, 99.9% or higher homology with the nucleotide sequence as shown in SEQ ID NO: 1; and (3) a nucleotide sequence with one or more, such as 1, 2, 3, 4, 5 nucleotide substitution, deletion or insertion, in the nucleotide sequence as shown in SEQ ID NO: 1. The clostridium butyricum provided by the invention can remarkably shorten the sleep latency period and improve the subthreshold dose hypnotic effect, and has a remarkable positive effect in the aspect of improving sleep. In addition, the clostridium butyricum provided by the invention has relatively high safety.

Genomically recoded organisms (GRO) are provided. In some forms, the GRO include no more than 25 genomic instances of the TGA codon. In some forms, genomically recoded organisms include two or more of single nucleotide substitutions, TAA insertions, and deletions to eliminate most or all of the TGA codons relative to a progenitor cell. In some forms, the GRO have a genome having stops codons, wherein the stop codons consisting of a single stop codon sequence. In some forms, the GRO provides translation termination only at the single stop codon sequence. Also provided are (i) a mutant release factor 2 (RF2) with reduced ability to terminate translation at UGA and / or a nucleic acid encoding the same and (ii) a mutant tRNATrp with reduced ability to decode UGA and / or a nucleic acid encoding the same, relative to a progenitor cell. GRO's expressing (i) and / or (ii) are also provided.

To provide a method for rescuing a termination codon through genetic reassignment using ACE-tRNA.SOLUTION: In certain embodiments, the invention provides a modified transfer RNA (tRNA) comprising a T-arm, a D-arm, an anticodon arm, and an acceptor arm, wherein the T-arm comprises a T-stem having a nucleotide that interacts with elongation factor 1 alpha 1 (EF1 alpha). EF1 alpha recruits aminoacyl-tRNAs to the ribosome and protects the tRNA from being deacylated. Rational nucleotide substitutions result in tuned tRNA: EF1 α interactions that enhance tRNA delivery to the ribosome and protection from de-acylation.SELECTED DRAWING: None

This technology provides a way to identify single nucleotide substitutions on microRNAs with sequence diversity at their 3' end in a short time and at low cost. [Solution] The method according to the embodiment is a method for detecting single nucleotide substitutions in a group of short RNAs having sequence diversity on the 3' end. The method includes: obtaining a target nucleic acid by adding an artificial sequence to the 5' end of a short RNA contained in the test nucleic acid; contacting the obtained target nucleic acid with a guide RNA having a sequence complementary to the target nucleic acid and single nucleotide polymorphism recognition sites and singularities at specific intervals, a genome editing enzyme corresponding to the guide RNA, and a label that becomes detectable upon activation of the genome editing enzyme; measuring the signal derived from the label; and determining the presence or absence of a single nucleotide substitution in the test nucleic acid from the intensity of the signal.

The present disclosure provides CRISPR / Cas9 ribonucleoprotein compositions comprising chemically modified CRISPR RNA (crRNA) guide and trans-acting CRISPR RNA (tracrRNA) components. Methods of using the disclosed CRISPR / Cas9 ribonucleoprotein compositions are also provided.

The present invention discloses a method and system for dynamically identifying risks of biotechnology products based on multi-indicator fusion, which relates to the field of biosafety management technology, including: S1, obtaining experimental data and environmental data from various stages of the life cycle of genetically modified crops, constructing a dynamic data set including molecular structure sequences, functional performance indicators and ecological impact factors, and extracting the mutation frequency of molecular structure sequences over time, abnormal fluctuations in functional performance indicators and dynamic change trends of ecological impact factors; S2, based on the mutation frequency of molecular structure sequences and abnormal fluctuations in functional performance indicators, extracting nucleotide substitution patterns and functional abnormality data sets of gene editing products in the research and development stage and the application stage, and determining the correlation pattern between nucleotide substitution patterns and functional abnormalities; this method and system for dynamically identifying risks of biotechnology products based on multi-indicator fusion realizes accurate prediction and dynamic management of ecological risks of genetically modified crops, and significantly improves the safety of environmental release.

The invention relates to the field of mycobacterium abscessus gene detection, and particularly discloses a nucleic acid probe group for MLST typing identification of a mycobacterium abscessus complex group and application of the nucleic acid probe group. The nucleic acid probe group comprises a target probe and a high GC balance probe; the target probe comprises a nucleotide sequence which contains substitution, deletion or insertion of 0-20 nucleotides or any combination of the substitution, deletion or insertion of the 0-20 nucleotides relative to the nucleotide sequence as shown in SEQ ID NO.1-188; the high GC balance probe comprises a nucleotide sequence as shown in SEQ ID NO.189, wherein the nucleotide sequence contains substitution, deletion or insertion of 0-20 nucleotides or any combination of the substitution, deletion or insertion of the 0-20 nucleotides. According to the invention, efficient enrichment and synchronous detection of MLST typing related targets are realized for the first time.

The invention provides a molecular marker of lactobacillus gasseri, the lactobacillus gasseri is preserved in China General Microbiological Culture Collection Center on February 7, 2023, and the preservation number is CGMCC No.26504. The molecular marker comprises one of the following nucleotide sequences: (1) a nucleotide sequence as shown in SEQ ID NO: 8, (2) a nucleotide sequence as shown in SEQ ID NO: 2, (3) a nucleotide sequence as shown in SEQ ID NO: 3, (4) a nucleotide sequence as shown in SEQ ID NO: 4, (5) a nucleotide sequence as shown in SEQ ID NO: 4, and (6) a nucleotide sequence as shown in SEQ ID NO: 4; (2) a nucleotide sequence having at least 97%, 98%, 99% or higher homology with the nucleotide sequence as shown in SEQ ID NO: 8; and (3) a nucleotide sequence with one or more nucleotide substitution, deletion or insertion in the nucleotide sequence as shown in SEQ ID NO: 8. By adopting the molecular marker disclosed by the invention, the target lactobacillus gasseri can be quickly, simply and conveniently screened; moreover, the target lactobacillus gasseri can be highly and specifically identified, and the interference of other strains is eliminated.

Provided are methods for detecting urogenital malignancies in dogs. In some embodiments, the methods include identifying a deletion or single nucleotide substitution within a BRAF gene and / or within a MAP2K1 gene present in or isolated from a biological sample from a. dog, wherein the presence of the deletion or single nucleotide substitution within the BRAF gene and / or within the MAP2K1 gene detects a urogenital malignancy, optionally transitional cell carcinoma / urothelial carcinoma, in the dog. In some embodiments, the deletion is within exon 12 of BRAF gene, optionally within the amino acid sequence KMLNVTAPTPQQL (SEQ ID NO: 3), and / or in within exon 2 or 3 of a M.AP2K1 gene, optionally within the amino acid sequence FLTQKQKVGE (SEQ ID NO: 4).

The present invention discloses a phage genome editing vector based on the CRISPR-Cas9 system and its editing method and application. red9 A plasmid containing the gene and sgRNA elements is constructed, and specific primers are designed for the editing site to obtain the pTarget plasmid; the donor DNA sequence is constructed into a vector to obtain the pEdit plasmid. After phage is infected with a host bacteriophage containing the pEdit plasmid, the pTarget plasmid is used to reverse screen for mutant phage, achieving efficient and rapid gene editing of the phage genome, including gene deletion mutations, single nucleotide substitution mutations, and insertion mutations. This method is simple to operate and has high gene editing efficiency. It has promoted the development of structural analysis and functional research of Staphylococcus aureus phage genomes and provided a new strategy for the modification of engineered phages, with broad application prospects and market value.

Genomically recoded organisms (GRO) are provided. In some forms, the GRO include no more than 25 genomic instances of the TGA codon. In some forms, genomically recoded organisms include two or more of single nucleotide substitutions, TAA insertions, and deletions to eliminate most or all of the TGA codons relative to a progenitor cell. In some forms, the GRO have a genome having stops codons, wherein the stop codons consisting of a single stop codon sequence. In some forms, the GRO provides translation termination only at the single stop codon sequence. Also provided are (i) a mutant release factor 2 (RF2) with reduced ability to terminate translation at UGA and / or a nucleic acid encoding the same and (ii) a mutant tRNATrp with reduced ability to decode UGA and / or a nucleic acid encoding the same, relative to a progenitor cell. GRO's expressing (i) and / or (ii) are also provided.

This invention provides a method for isolating target microorganisms, which are rare biospheres, from environmental microbial samples. [Solution] A method for isolating target microorganisms, which are rare biospheres, from an environmental microbial sample, comprising the steps of: preparing donor DNA, which is nucleic acid around the sequence of an antibiotic susceptibility gene, to perform a single nucleotide substitution on the antibiotic susceptibility gene of the target microorganism; introducing the donor DNA into the environmental microbial sample and performing a single nucleotide substitution on the antibiotic binding site of the target microorganism; accumulating and culturing the environmental microbial sample in a liquid medium containing the antibiotic; and separating the microorganism after accumulation and culturing by the agar plate method or the ultradilution method.

This invention discloses a highly efficient and scarless method for editing the genome of myxobacteria and its applications. This invention combines the ISDra2 TnpB cleavage system of *Gastrococcus radiodurans* with the MxRedET homologous recombination system of myxobacteria to form a one-step scarless editing platform (MxDIRECT). ISDra2 is responsible for precisely locating and cleaving the target DNA, and MxRedET then efficiently integrates or replaces fragments at the cleavage site, enabling gene knockout, insertion, and replacement operations. This method increases the editing efficiency of myxobacteria from 1% to 100%, shortens the cycle from 3-6 months to 5-7 days, and allows for one-step two-site scarless editing through a two-site design. A single cleavage can knock out up to approximately 80kb of DNA, while two cleavages can knock out up to 200kb; and single nucleotide substitutions can be performed at any location, achieving precise base editing. This technology provides an efficient and precise tool for modifying the functional genes and metabolic pathways of myxobacterial chassis cells, significantly advancing synthetic biology research.