39 results about "Metagenomic dna" patented technology

The invention relates to a new glycosyl hydrolases with an amyloltic activity and nucleic acids coding for said gylcosyl hydrolases, A PCR-based method for identifying and preparing new gylcosyl hydrolases from metagenome DNA and several possible technical uses for such glycosyl hydrolases with an amylolytic activity. Washing and cleaning products containing such enzymes, and methods and possible uses corresponding thereto are particularly interesting.

The invention discloses a method to sieve scale microbe alkali proof relative gene in microbe engineering domain, which is characterized by the following: collecting typical basic environment soil sample; rough-extracting; purifying; getting macro-genom DNA of soil sample; electrization-inverting bacillus coli defect strain E with connecting product between macro-genom DNA part enzyme cutting fragment and carrier; sieving; getting positive colony; proceeding sub-clone and functional appraise for alkali proof related gene of positive clone. This invention can be used to breeding of alkali proof and molecule of microbe.

The invention discloses a key microbial functional genome detection method in a pepper peeling process. According to the key microbial functional genome detection method in the pepper peeling process, from the perspective of micro-ecosystem, pepper epidermis microorganisms in the peeling process are taken as an object; metagenome DNA of pepper epidermis bacteria is extracted and purified and a variable region sequence is amplified, and then the variable region sequence of the epidermis bacteria is read with the application of a high-throughput sequencing technology; by virtue of a bioinformatics platform, the sequence is subjected to quality control and species annotation and function prediction are completed, and on the basis, a microorganism species, which plays a vital role in the pepper peeling process, in fresh fruit epidermis and a core metabolic pathway thereof are comprehensively and objectively revealed depending on a statistical method. The detection method provided by the invention simplifies experimental operations and improves a sequencing efficiency, so that the development of metagenomics is greatly promoted.

The invention discloses a method for real-time fluorescence quantification PCR (qPCR) detection of denitrifying microorganisms in aquaculture environment sediments, and belongs to the technical field of environmental microbial ecology. The method comprises the following contents: metagenome DNA extraction and concentration determination of sediment samples in various stations, PCR amplification of nirK and nirS genes, construction of a standard plasmid, preparation of a standard curve, qPCR amplification and calculation of the number of denitrifying microorganisms in the samples according to a qPCR result in accordance with the standard curve. The method disclosed by the invention can be used for determining the number of the denitrifying microorganisms in an aquaculture environment through quantitative analysis on two genes, namely nirK and nirS, so as to overcome the shortcoming that 16S rRNA gene cannot be used in the quantification of the denitrifying microorganisms as well as the shortcoming that the prior art, which focuses on some functional gene only, is incomplete in quantification; and the provided method for detecting the number of the denitrifying microorganisms in the aquaculture environment is good in specificity, high in repeatability and high in accuracy. According to the method disclosed by the invention, logarithm values of nirK and nirS gene copy numbers keep a good linear relation with Ct value of a luminescence threshold, and the regression coefficient of the standard curve is above 0.99.

A method for selectively obtaining a natural variant of an enzyme having activity includes (1) a step of detecting an ORF sequence of a protein having enzyme activity from a genome database including base sequences of metagenomic DNA of environmental microbiota; (2) a step of obtaining at least one PCR clone including the ORF sequence having a full length, a partial sequence of the ORF sequence, or a base sequence encoding an amino acid sequence which is formed by deletion, substitution, or addition of at least one amino acid residue in an amino acid sequence encoded by the ORF sequence, by performing PCR cloning on at least one metagenomic DNA of the environmental microbiota by using a primer designed based on the ORF sequence; (3) a step of determining a base sequence and an amino acid sequence which is encoded by the base sequence for each PCR clone obtained in the step (2); and (4) a step of selecting a natural variant of an enzyme having activity by measuring enzyme activity of proteins encoded by each PCR clone obtained in the step (2).

The present invention discloses an extraction method for metagenome, and a method of probe hybridization-bead capture is used to reduce the host DNA content in a host metagenomic DNA sample. According to the method, the Alu repeat sequence of the host DNA sequences and conserved sequence at both ends thereof are used as templates to design one-way or two-way probes, wherein the 5 'end of each probe is modified with biotin, streptavidin coated beads are used to capture host DNA hybridized with probes, in order to achieve aims of weakening host DNA background in a metagenome, and enhancing sequencing data efficiency.

The present invention provides an oral throat swab bacterial metagenome DNA extraction method, which comprises five steps such as oral throat swab pretreatment, bacterial lysis, DNA separation and hybrid protein removing, adsorption of DNA through a silica gel membrane purification column and impurity removal, and elution of DNA so as to extract the oral throat swab bacterial genome DNA. According to the present invention, with the special sample treatment method, the yield can be substantially increased; the combination of ultrasonic wave crack cleavage and enzyme cleavage is used, and the appropriate wall breaking time and the mild conditions are used, such that the gram-negative bacteria, the gram-positive bacteria and other bacteria are subjected to appropriate wall breaking, and the breaking of the genomic DNA due to the excessive strong physical factors can not be produced; and the oral throat swab bacterial metagenome DNA can be extracted only in 1.5 h, and the bacterial metagenome yield is high.