5 results about "Ribosomal DNA" patented technology

This disclosure generally relates to fusion proteins comprising TAL Arrays targeting a repetitive element and a transposase domain comprising amino terminal deletions, as well as dual cysteine rich domains (CRD), for targeting site-specific transposition into ribosomal DNA (rDNA) repeats or LINE1 repetitive elements, polynucleotides and vectors encoding the fusion proteins, and methods of use therefor.

The present invention relates to a method for detecting and identifying a microorganism in a biological sample by a nucleic acid amplification reaction, said method comprising the steps of: providing a biological sample suspected of comprising a microorganism, and optionally isolating nucleic acids from said biological sample; b) providing a reaction container comprising a nucleic acid amplification reaction mixture comprising an aliquot of said biological sample or nucleic acids isolated therefrom, at least one pair of broad-taxonomic-range amplification primers for amplifying a microbial ribosomal DNA (rDNA) internal transcribed spacer (ITS) region from the, optionally isolated, nucleic acids in said sample and for generating an rDNA ITS amplicon, at least one taxon-specific detection probe for detecting a taxon-specific sequence in said rDNA ITS region, a DNA intercalating dye for recording a high resolution melting curve for said rDNA ITS amplicon; performing a nucleic acid amplification reaction on the mixture to generate said rDNA ITS amplicon; recording an hrMC for said rDNA ITS amplicon; comparing the high resolution melting curve recorded with a database comprising hrMCs of reference amplicons generated from reference microbial species of known taxonomic identity using the same set of broad-taxonomic-range amplification primers, to thereby obtain a first taxonomic identity indicator of a microorganism present in said sample; detecting hybridization between said at least one taxon-specific detection probe and said rDNA ITS amplicon to thereby obtain a second taxonomic identity indicator of a microorganism present in said sample, and identifying the microorganism present in said sample at species level based on said first and second taxonomic identity indicator, wherein steps c), d), and f) are carried out in a closed reaction container.

This invention relates to LAMP primer sets, kits, and applications for detecting *Pseudomonas aeruginosa*, belonging to the field of molecular biology. Based on the conserved internal transcribed spacer (ITS) region of the ribosomal DNA of *Pseudomonas aeruginosa*, this invention designs in vitro DNA amplification primers. Through color reactions and electrophoretic banding after in vitro amplification, a simple and rapid method for detecting *Pseudomonas aeruginosa* is established. This method solves some technical problems existing in the early diagnosis of peony yellow spot disease based on morphological characteristics combined with molecular sequence identification. Compared with existing technologies, the LAMP detection technology for *Pseudomonas aeruginosa* established in this invention is rapid, efficient, low-cost, highly sensitive, highly specific, and convenient for product detection, providing a theoretical basis for the early diagnosis and timely control of peony yellow spot disease in production.

The invention belongs to the technical field of biology, and provides a method for identifying a ribosome DNA autonomous replication sequence and application. A ribosome DNA sequence of a target bacterium is obtained by adopting a three-section amplification theory, free plasmids are constructed, the target bacterium is transformed, and the ribosome DNA autonomous replication sequence of the target bacterium is screened by determining the transformation efficiency and evaluating the existence form and loss rate of the plasmids. According to the invention, the DNA autonomous replication sequence is identified from ribosome DNA, which is easier than that from genomes, especially for non-model organisms. The technology belongs to an enabling technology, and solves the problems of high complexity and low success rate of the prior art. In addition, a three-section amplification strategy is adopted to identify the independent replication sequence of the ribosome DNA, so that the method is simpler, more convenient and quicker than the existing method, the identification of the independent replication sequence of multiple groups of species is facilitated, genetic tools are perfected, and the technical development and application development of microorganisms are assisted.

The present disclosure provides, in part, recombinant, mutant Bxb1 integrases which are able to target pseudo-aft sites which are newly discovered "safe harbor" genomic locations, e.g., found in ribosomal DNA (rDNA), and method of engineering nucleic acids and genomic insertion of host cells using the same.