39 results about "Direct repeat" patented technology

The embodiment of the invention discloses a method for identifying clustered regularly interspaces short palindromic repeats (CRISPR). The method comprises the following steps: according to a DR (Direct Repeats) template in a source file, determining a first-generation CRISPR; after the missed DR in an undetermined spacer sequence between two pieces of adjacent first-generation CRISPR is added, determining a second-generation CRISPR; determining the CRISPR of which the SPACER similarity is lower than a preset threshold value in the second-generation CRISPR as a third-generation CRISPR; and determining the third-generation CRISPR of which two ends are provided with the DR as a four-generation CRISPR. The embodiment of the method can reduce misinformation or ignore the cut-off DR so as to improve the identification accuracy and comprehensiveness of the CRISPR.

The invention discloses an integrative plasmid pOPHI and a resistance screening marker-free self-luminescent mycobacterium. The integrative plasmid pOPHI comprises a promoter, an enzyme gene (LuxCDABE) needed for luminescence, a fusion gene of an integrase gene (Int) and a resistance screening gene, direct repeat sequences DifR and DifL positioned on two ends of the fusion gene, a phage integration site (attP) and a replicon. After the integrative plasmid pOPHI is transferred into the mycobacteria, a photobacterium of which the resistance gene and the integrase gene are lost is screened out by subculturing, and the photobacterium is the resistance screening marker-free self-luminescent mycobacterium. Whether the resistance screening marker-free self-luminescent mycobacterium obtained by construction is dead or not is judged according to whether the bacterium is luminescent or not, the mycobacterium can be used for a plurality of detections of experiments, and credible experimental data can be quickly obtained.

The invention discloses a method of constructing selection-marker-free self-luminescent mycobacterium abscessus and establishment of a corresponding in-vitro high-flux medicine-screening model. During the construction process, a key factor is the construction of a recombinant plasmid pOPAI1B, which contains a fused gene including a luminescent required enzyme gene LuxCDABE, a resistance screening gene Apr and an integrase gene Int; direct repeat sequences DifR and DifL at the two ends of the fused gene respectively; and a phage integration site attP and a replicon. The recombinant plasmid pOPAI1B is transformed into the mycobacterium abscessus and then is screened to obtain the selection-marker-free self-luminescent mycobacterium abscessus of which a resistance gene and the integrase gene are both lost. The self-luminescent mycobacterium abscessus can screen drugs in a high flux, can determine sterilizing or bacteria-resistant effects of the drug by means of detection of luminescent situation of bacteria, can be used for in-vitro drug activity detection, is free of any substrates, can continuously detect the same sample, is high in sensitivity and is strong in repeatability.

The invention discloses an oligonucleotide composition and a method for detecting hepatitis B virus (HBV) rcDNA (Relaxed Circular deoxyribonucleic acid) and/ or cccDNA (Covalently Closed Circular DNA), a kit and application of the oligonucleotide composition. The oligonucleotide composition comprises first oligonucleotide, second oligonucleotide, or oligonucleotide complementary with the sequenceof the first oligonucleotide and oligonucleotide complementary with the sequence of the second oligonucleotide; the first oligonucleotide is specifically combined with at least one part of continuoussequence of the first sequence, and the second oligonucleotide is specifically combined with at least one part of continuous sequence of the second sequence; in addition, a distance between two continuous sequences is 30-350 nt; the first sequence is a sequence between DR (Direct Repeat) 2-DR1 of the HBV DR sequence of a cross-notch area which contains cccDNA or rcDNA, and the second sequence is asequence after the fifth basic group of the DR1. According to the oligonucleotide composition, the level of HBV rcDNA and/ or cccDNA can be effectively detected without being affected by integrated HBV DNA.

A probe combination for detecting cancer includes one or more sets of partial hepatitis B virus (HBV) targeting probes. When sequences of each of the sets of partial HBV targeting probes are aligned, an overall sequence of the aligned set of probes matches a reference sequence of a genome of a HBV genotype or a direct repeat (DR) region on the genome. In the aligned set of probes, each of the probes overlap with one or two adjacent probes by a portion of a length of the probe. The probe combination may further includes one or more sets of hotspot gene targeting probes targeting cancer hotspot genes such as CTNNB1, TERT, and TP53 genes, one or more sets of exogenous gene targeting probes targeting portions of a lambda phage genome, and endogenous gene targeting probes targeting endogenous genes such as GAPDH and GdX genes.

The invention relates to a tobacco chloroplast genome recombination hotspot short-sequence NtcpRH23. According to the invention, the obtaining of chloroplast-converted tobacco material is employed as a basis, and a discovery is that: a segment of 23bp short-sequence NtcpRH23 on the constructed carrier can be subject to high-frequency recombinations with upstream direct repeats. According to PCR verifications, recombination events occur in four obtained conversion materials, and tobacco chloroplast genome recombinants are produced. The discovery of the segment of sequence assists in providing a novel molecular biological tool for the construction of chloroplast conversion novel carrier, and for the researches of relevant biological hot problems of chloroplast genome copying, chloroplast genome recombination, and chloroplast genome restoring mechanism.

The present invention relates to enhanced Sleeping Beauty-type transposons and methods of transposition. In particular, the invention relates to a polynucleotide comprising a cargo nucleic acid flanked by a left and a right inverted repeat/direct repeat (IR/DR), wherein IR/DRs, having specific sequences, are recognized by a Sleeping Beauty transposase protein and the polynucleotide is capable of integrating into the DNA of a cell. The invention also relates to a kit for transposing a nucleic acid comprising said polynucleotide as well as to further components such as co-factors of transposition capable of depleting a component of the FACT (facilitates chromatin transcription) complex, namely, SSRP1 and/or SUPT16H/SPT16,or an inhibitor of cathepsin selected from the group comprising H,S,V,and L; or a cofactor capable of depleting or inhibiting HSP90; or a factor temporally arresting cells cell cycle in cell cycle phase G0/G1, G1/S, or G2/M; or a factor inhibiting the ubiquitination ofPCNA,or cells wherein these components have been knocked down or inhibited,or the cell cyle arrested in any of said stages. Alternatively or additionally, the kit may comprise as a co-factor of transposition an agent capable of increasing concentration and/or signaling of ATR or a cell wherein concentrationand/or signaling of ATR are increased. The invention further provides methods using said transposon polynucleotide as well as host cells and pharmaceutical compositions. It also relates to use of said co-factors of transposition or specific cells for enhancing transposition efficiencies, e.g., for preparing genetically modified nucleic acids or cells.

The invention discloses a method for introducing site-specific mutagenesis rapidly and efficiently. The method provided by the invention can realize the introduction of site-specific mutagenesis by using an integrated plasmid YIplac211 as the carrier, selecting URA3 gene carried by the integrated plasmid as selection marker, and carrying out homologous recombination of direct repeats twice. According to the invention, no exogenous gene remains during the introduction of site-specific mutagenesis so as to ensure the safety of strains. Additionally, the method provided by the invention can also use other industrial strains besides yeast.

It is intended to provide an assay for the presence, absence or amount of a nucleic-acid fragment having a certain nucleotide sequence, for example, a polyA length, a difference in the number of repetition of a direct repeat sequence (e.g., microsatellite), single nucleotide substitution (or single nucleotide polymorphism), and nucleotide sequence insertion or deletion, and to provide a genetic testing using the same. The present invention relates to a nucleotide analysis method, comprising: hybridizing at least two probes to a nucleic-acid fragment; ligating the at least two probes using ligase; exchanging, to ATP, pyrophosphoric acid produced through the ligation reaction; and detecting chemiluminescence reaction dependent on the ATP.