53 results about "Homologous Sequences" patented technology

The application discloses a PCR primer pair and a molecular marker probe for distinguishing between barnyard grass and rice, a detection kit and application thereof. After align analysis on chloroplast genome sequences of the barnyard grass and the rice, it is found that a difference site exists in a 91 kb region, sequences of the upstream and downstream of the difference site are taken as reference sequences to select homologous sequences to design primers for PCR amplification, and the amplification result proves that the difference site fragment obtained by chloroplast genome sequence analysis exists and can be amplified and identified subsequently; the PCR amplification product is sequenced, and the sequencing result is compared to find that a 12 bp insertion site exists in the site and can separate the barnyard grass from the rice, a colloidal gold probe is designed according to the insertion site, and a RPA detection kit and a detection method are established, and the RPA detection kit is used to detect the barnyard grass and the rice, and the barnyard grass and the rice can be accurately distinguished.

The invention relates to the technical field of gene engineering, and provides application of polynucleotide, protein and a biological material in regulation and control of plant nutrient substance storage organ development. The polynucleotide has a sequence as shown in SEQ ID NO: 1; or a complementary sequence, a degenerate sequence or a homologous sequence thereof; or polynucleotide hybridized with the nucleotide sequence as shown in SEQ ID NO: 1 under strict conditions or a complementary sequence of the polynucleotide. The polynucleotide is related to the development of plant nutrient substance storage organs, promotes the development of the plant nutrient substance storage organs through overexpression, and can be used for cultivating plant varieties.

A homologous recombination method of excising a specific DNA sequence from a DNA molecule. The method includes providing the DNA molecule including a pair of target sequences that are a target of a site-specific recombination enzyme and a pair of homologous sequences that are DNA sequences sharing homology with each other. The pair of target sequences are oriented in different directions from each other and arranged on opposite sides of the specific DNA sequence. The pair of homologous sequences are arranged on opposite sides of the specific DNA sequence at positions further outside of the pair of target sequences. The method further includes inducing homologous recombination between the pair of homologous sequences by action of the site-specific recombination enzyme on the DNA molecule so that the specific DNA sequence and the pair of target sequences are excised from the DNA molecule.

The invention discloses a Chinese cabbage clubroot-resistant gene BrRLP31 and application of the Chinese cabbage clubroot-resistant gene BrRLP31. The nucleotide sequence of the Chinese cabbage clubroot-resistant gene BrRLP31 is as shown in SEQ ID No. 1. Functional verification results show that after the expression of the endogenous BrRLP31 of the Chinese cabbage is inhibited by utilizing a VIGS technology, the resistance of the plant to clubroot is obviously reduced, and the disease index and the root swelling severity of the plant are obviously higher than those of a control material. On the other hand, the transgenic flowering cabbage plant over-expressed by the BrRLP31 is constructed by an agrobacterium-mediated method, and disease resistance identification proves that the heterologous over-expression of the BrRLP31 obviously improves the resistance of the plant to plasmodiophora brassicae. The results show that the BrRLP31 gene plays a key positive regulation role in the clubroot resistance process of the Chinese cabbage, the disease resistance of cruciferae crops and other horticultural plants is improved by utilizing the gene and the homologous sequence thereof, and the gene and the homologous sequence thereof have important application value.

The invention discloses a method for detecting wheat differential transposon elements based on genome scanning, and belongs to the technical field of biology. The core of the invention focuses on a core technology system of crop genome TE difference detection and an application scene thereof, and comprises a detection method and a verification method. A homologous sequence distinguishing method based on an A / B / D subgenome SNP tag is protected, and the method comprises flanking region SNP matching rate calculation, a sliding window correction algorithm, a TE polymorphism extraction program and the like. The detection method provided by the invention effectively solves the problems of low efficiency of whole genome structure variation analysis, low detection specificity caused by polyploidy homologous sequence interference and high false positive of a single PCR primer pair verification system.

The invention discloses a leaf and branch compost cutinase link and application thereof, and belongs to the technical field of enzyme molecule construction. The leaf-branch compost cutinase kink is obtained by adjusting the connection sequence of fragments in leaf-branch compost cutinase, and comprises a fragment II, a connecting peptide fragment I, a fragment I, a connecting peptide fragment II and a fragment III which are connected in sequence from the N end to the C end; wherein the fragment I, the fragment II and the fragment III respectively correspond to 37th to 79th amino acids, 81st to 143rd amino acids and 150th to 293rd amino acids or homologous sequences thereof of the leaf branch compost cutinase. According to the method, protein topology engineering and an artificial intelligence assisted protein design technology are organically combined, knot topology transformation is performed on the leaf branch compost cutinase, sequence optimization is performed on a connecting peptide fragment of the knot, and the leaf branch compost cutinase knot with good biological activity is obtained; the expression quantity and the stability of the leaf and branch compost cutinase kink are improved, the problem of kinetic barriers existing in protein kink combination is solved, and the method has important application value.

The present application relates to a multiplex PCR primer probe combination and kit for human red blood cell RHD gene typing detection, and belongs to the field of biomedical clinical molecular detection. The multiplex PCR primer probe combination for human red blood cell RHD gene typing detection comprises 19 pairs of specific primers and corresponding probes distributed in 8 reaction wells. The present application also provides a kit containing the multiplex PCR primer probe combination for human red blood cell RHD gene typing detection. The RHD gene variant site specific primers and probes designed by ARMS combined with homologous sequence specific base method are more specific and accurate; all high frequency RHD gene variants of Chinese population can be detected by one experiment, including DEL type (RHD*DEL1, c.1227G>A), weak D15 type (RHD*15), partial RHD*DVI.3 type (RHD*DVI.3), RHD deletion type D negative (RHD*01N.01) and other common D negative phenotypes of Chinese population, RHD*D-CE(2-9)-D (RHD*01N.03) and RHD*01N.16 (c.711delC), and normal RHD positive types (RHD*01) can also be detected.

The invention discloses siRNA (small interfering Ribonucleic Acid) of targeted bone morphogenetic protein 2, modified siRNA and application of the siRNA. According to the invention, siRNA drug design is carried out by taking the BMP2 gene as a target gene to obtain a series of siRNAs, siRNAs capable of effectively silencing the mRNA expression level of the BMP2 gene are screened from the siRNAs, and part of siRNA sequences can even silence the mRNA level of the BMP2 gene to about 25%. The siRNA sequence is further modified, wherein phosphorylation or phosphorylation analogue modification, skeleton modification, sugar ring modification and basic group modification are carried out on the 5'terminal of the sequence, or cholesterol molecule modification is carried out on the end group of the siRNA. The siRNA sequence is further subjected to cholesterol modification to obtain a human-mouse homologous sequence of the targeted BMP2 gene, and the in-vivo silencing activity of the BMP2 target spot is further researched in a mouse experimental model subsequently.

The invention belongs to the technical field of molecular biological detection, and relates to a polypeptide for detecting asymptomatic infection of plasmodium vivax malaria and application. The polypeptide for detecting the asymptomatic infection of the malaria of the plasmodium vivax is an amino acid sequence as shown in SEQ ID No.1, or a homologous sequence which is subjected to substitution, deletion or insertion of one or more amino acids or has the sequence similarity greater than 75%. The polypeptide has an amino acid sequence as shown in SEQ ID No. 1. The polypeptide SEQ ID No.1 in the invention specifically detects asymptomatic tertian malaria infection, and can effectively deal with tertian malaria input threats in the Burmese border area. Meanwhile, the polypeptide can be coated in a detection tool of a rapid detection kit, and the tool has the characteristics of portability, high efficiency, low cost and the like, and is suitable for field sample detection in a malaria prevalence area.

The invention discloses a synthetic protein design method based on protein feature engineering and a diffusion model, which comprises the following steps: firstly, searching a homologous sequence of a query sequence in a non-redundant NR database by using a P-BLAST tool and taking a target protein sequence as the query sequence; carrying out ProtParam-7 and ProtFactor-5 feature engineering on the screened homologous sequences, and carrying out ProtParam-7 and ProtFactor-5 feature engineering on the screened homologous sequences; taking data represented by the ProtParam-7 and ProtFactor-5 characteristics as original proteins, and inputting the original proteins into the diffusion model; performing multi-dimensional screening on the generated candidate proteins; evaluating the functional activity and the structural stability of the candidate protein by adopting a wet experiment, and taking the protein meeting the condition as a final generation result. According to the method, protein sequence feature extraction and diffusion model generation are combined, and multi-dimensional screening and experimental verification are carried out to ensure that the generated protein is close to the target protein in structure and function.

The invention discloses a recombinant peanut allergen fusion protein and application thereof, the amino acid sequence of the recombinant peanut allergen fusion protein is as shown in SEQ ID NO.1, and the corresponding coding nucleotide sequence of the recombinant peanut allergen fusion protein is as shown in SEQ ID NO.2. The recombinant peanut allergen fusion protein comprises an Ara h 8 protein 1-157aa sequence fragment, an Ara h 9 protein 1-116aa sequence fragment and an Ara h 2 protein 22-172aa sequence fragment, the three proteins are sequentially connected through a Linker sequence to obtain the peanut allergen fusion protein, and a recombinant fusion expression method of the fusion protein and a detection kit based on the protein are provided. The fusion protein and the kit thereof can be applied to peanut desensitization treatment, can also improve the positive detection rate in immunodiagnosis application and eliminate cross interference caused by homologous sequences of other allergens, and can improve the performance of a peanut allergy diagnostic reagent as a raw material.

The present invention relates to a subtilisin BPN' variant or homologue thereof comprising new mutations compared to subtilisin BPN' as shown in SEQ ID NO: 2 or a homologous sequence thereof. Such mutations can occur at amino acid positions selected from the group consisting of L96, D99, A223 and S224. The present invention also relates to a method for the enzymatic synthesis of a peptide by coupling of peptide fragments, wherein said coupling is catalysed by said subtilisin BPN' variant or homologue thereof.

A method for producing a SARS-CoV-2 virus-like particle-based protein subunit vaccine. The method comprises creating a fusion protein by combining a DNA sequence encoding an iLOV protein with a DNA sequence encoding a peptide linker and a cleavage site for enterokinase protease and a DNA sequence encoding a Receptor Binding Domain (RBD) of the SARS-CoV-2 viral spike protein. The RBD protein is attached to a “Spy Tag” peptide. The peptide linker cleavage site DNA sequence is between the iLOV protein DNA sequence and either the SARS-CoV-2 viral protein DNA sequence or the “Spy Tag” peptide DNA sequence. The DNA sequence encoding the fusion protein is introduced into a P. pastoris host to form transformants. At least one optimal recombinant is identified from the transformants using fluorescence to detect optimal expression levels of the SARS-CoV-2 viral protein. The SARS-CoV-2 viral protein is isolated from the fusion protein by cleaving the iLOV protein and linker sequences from the target protein. The RBD sequence can be mutated to represent RBD variants or homologous sequences or improve expression and alter its glycosylation pattern. The RBD sequence can belong to any virus within the coronavirus family.

The invention belongs to the technical field of gene editing, and particularly discloses a Crimson fluorescent protein reporting system and application thereof in gene editing efficiency multi-scene evaluation. According to the Crimson fluorescent protein reporting system, an equidirectional repetitive sequence which can be targeted by sgAlb and contains a termination codon is introduced into a Crimson gene sequence, a frame-shift mutant fluorescent reporting module is successfully constructed, and the module can recover fluorescent expression through an SSA (homologous sequence annealing repair) repair mechanism after being cut in a Cas9 / sgAlb system. The developed Crimson report cell system can be used for accurately evaluating the stability and editing efficiency of different sgRNA skeletons; the method is suitable for optimizing a virus transduction system and screening a guiding aid for efficient gene delivery; plasmid pollution in a virus vector system can be rapidly detected, and accurate distinguishing of a functional RNP compound and residual plasmid DNA is realized by comparing the proportion of Crimson positive cells before and after DNase I treatment and heat treatment; and small molecular compounds for promoting homologous directional repair can be efficiently screened.

The application relates to the technical field of bioinformatics, and discloses a urease catalytic performance control method, device, equipment and storage medium. The method comprises the following steps: obtaining a wild-type amino acid sequence of a target urease and a three-dimensional structure corresponding to the wild-type amino acid sequence; mapping the amino acid sequence into a sequence feature unit, and mapping a local structure feature extracted from the three-dimensional structure into a structure feature unit; jointly encoding the sequence feature unit and the structure feature unit by using a decoupling multi-head cross attention module to obtain original logic values; constructing a multiple sequence alignment file based on homologous sequences, counting the amino acid frequency distribution of each residue position, and generating evolution logic values; determining an initial candidate mutant set; constructing an initial prediction model and a target prediction model; and predicting the initial candidate mutant set based on the target prediction model to obtain a target mutant with target catalytic performance. The scheme improves the control stability of urease catalytic performance.

This invention belongs to the field of enzyme catalysis and bioengineering technology, and relates to a PET hydrolase, Marine0715, and its applications. The enzyme is a GFP11-Marine0715 fusion protein with a GFP11 fusion fragment at its N-terminus, and its amino acid sequence is shown in SEQ ID NO.2 of the sequence listing. This invention also provides the nucleic acid encoding this enzyme, a recombinant vector carrying the encoding nucleic acid, and a recombinant expression strain. The Marine0715 provided by this invention is derived from a marine metagenomic genome, with a highest homology sequence identity of less than 90%, possessing novelty and value as a new enzyme engineering backbone; it expands the existing PET hydrolase sequence resource library.

The application discloses a primer pair, a method and a kit for detecting TUBB8 gene mutation. The primer pair is shown in SEQ ID NO:1 and SEQ ID NO:2, and a long fragment product covering a complete gene locus of the TUBB8 gene is specifically amplified. The method comprises the steps of extracting genomic DNA, long fragment PCR amplification, constructing a sequencing library, long read sequencing and gene variation analysis. The primer design of the application effectively avoids homologous sequence interference, solves the problem of high false positive rate in the prior art, has comprehensive detection range, can find point mutation, insertion and deletion, structural variation and regulatory region variation at one time, supports an efficient clinical sequential diagnosis path of "targeting TUBB8 first and then whole exon sequencing", and has significant accuracy, economy and clinical application value.

The invention discloses a high-fidelity Cas9 homolog based on structure orientation and a screening method and application thereof, and belongs to the technical field of gene editing. According to the method, cross-database searching is carried out, a sequence comparison tool is used for finding a homologous sequence, duplicate removal is carried out, three candidate enzymes are effectively recognized from 24 and 544 Cas9 homologues by combining the sequence and three-dimensional structure evaluation, engineering variants with improved editing efficiency are obtained through PAM recognition area replacement and rational design, and the eSeqCas9 editing efficiency reaches 82.9%. The high-fidelity Cas9 homologue and the engineered variant thereof have good performance in a base editing system, the editing efficiency of A-to-G and C-to-T is considerable, and whole genome off-target detection shows that the Cas9 homologue and the engineered variant thereof have higher specificity. A new thought is provided for discovering the high-fidelity Cas9 homologue, and when the Cas9 homologue is applied to a gene editing tool, accuracy and safety can be improved, off-target can be reduced, and development of the fields of gene therapy, biological breeding and the like can be promoted.

The present invention relates to a method for the enzymatic synthesis of (oligo)peptides comprising coupling (a) a (oligo)peptide C-terminal ester or thioester and (b) a (oligo)peptide nucleophile having an N-terminal unprotected amine, wherein the coupling is performed in a fluid comprising water, and wherein the coupling is catalyzed by a subtilisin BPN' variant or a homologue thereof comprising the following mutations compared to subtilisin BPN' as set forth in SEQUENCE ID NO: 2: deletion of the amino acids corresponding to positions 75-83; a mutation at the amino acid position corresponding to S221 which is S221C or S221 selenocysteine; preferably a mutation at the amino acid position corresponding to P225, wherein the amino acid positions are defined according to the sequence of subtilisin BPN' as set forth in SEQUENCE ID NO: 2. In addition, the present invention relates to enzymes suitable for use as catalysts in the method of the present invention.

The invention relates to the cross technical field of bioinformatics and epigenetics, and particularly discloses a method for rapidly identifying plant imprinting genes based on multi-species sequence conservation.The method comprises the steps that a whole genome gene set of target plant species is obtained; selecting reference plant species with different genetic relationships with the target plant species, obtaining an optimal homologous gene of each gene of the target plant species in the reference plant species, and constructing a multi-species homologous sequence set; performing multi-sequence comparison on the multi-species homologous sequence set, and calculating a conservative score of each target gene based on a comparison result; and sorting and screening the target genes based on the conservative scores, and preliminarily identifying the genes with the conservative scores higher than a set threshold as candidate plant imprinting genes. According to the method, high-throughput and low-cost screening and prediction of potential plant imprinting genes are realized by utilizing the characteristic that the plant imprinting genes have higher conservative property in an evolution process and calculating cross-species conservative property scores of the genes.

The application belongs to the technical field of biology, and particularly relates to a grass carp reovirus (GCRV-Ⅱ type) subunit recombinant vaccine, a preparation method and application. 274‑367 , VP5 105‑336 , NS38 143‑338 sequence fragments, and the three sequence fragments are recombined and expressed in a prokaryotic expression vector to prepare the subunit recombinant vaccine. The recombinant vaccine has high safety, strong activity and broad spectrum, and has good application potential.

This invention discloses a method and its application for predicting the trans-splicing activity of fragmented inpeptides. The method takes the amino acid sequence of the fragmented inpeptide as input, extracts 1280-dimensional residue-level sequence embedding features using the ESM-2 protein language model, and uses AlphaFold2 for high-precision three-dimensional structure prediction to obtain the spatial coordinates of the residues. A graph structure is constructed with residues as nodes and residue pairs with a Cα atomic distance of less than or equal to 8 Å as edges. Node features include sequence embedding and chain affiliation information, while edge features include Cα distance and cross-chain connection identifiers. The graph structure is input into the DeepIntein deep learning model based on a graph attention network, and the splicing activity prediction probability is output after multi-layer graph attention convolution, global pooling, and fully connected layers. The prediction throughput of this invention is more than three orders of magnitude higher than that of the traditional Western blotting method, breaking through the limitations of homology sequence exploration space.

This invention relates to the field of genetic engineering technology, specifically to a recombinant plasmid, its construction method, and its application in enhancing the synthesis of pseudocladocerin from Nostoc flagelliforme. The recombinant plasmid includes a vector plasmid, and sgRNA target sequences, HR homologous sequences, psbA promoter sequences, and other components linked to the vector plasmid. ScyR Regulatory genes. This recombinant plasmid can be used in black moss. MAAs Overexpression of key regulatory genes at gene cluster sites significantly enhanced the pseudocranophane synthesis capacity of Nostoc commune. The Nostoc mutant obtained by transforming this recombinant plasmid into Nostoc commune exhibited a pseudocranophane content 2.4 times higher than that of wild-type Nostoc commune after 15 days of culture.

The invention discloses a bird and animal keratinized tissue mitochondrial DNA assembly method. The method comprises the following steps: acquiring second-generation sequencing data and cutting a low-quality sequence to generate a clean file; carrying out comparison until a mitochondrial reference genome is obtained, and screening a homologous sequence, so as to obtain unclassifid.sam; analyzing terminal base damage, counting significant damage and determining a parameter N; the SAM is converted into a mapped.fq.gz file; generating a pre-processing file based on N-substituted forward data 5'end front N basic groups and reverse data 3 'end front N basic groups; and iteratively assembling and outputting a complete mitochondrial DNA sequence by using a preset k value, thereby improving the speed and integrity.

The application discloses an anti-HpFucT protein monoclonal antibody and products and applications thereof, and belongs to the biomedical technical field.The HCDR1, HCDR2 and HCDR3 of the antibody heavy chain variable region are respectively shown as SEQ ID No.1-3 or a homologous sequence thereof, the LCDR1, LCDR2 and LCDR3 of the light chain variable region are respectively shown as SEQ ID No.6-8 or a homologous sequence thereof, or are respectively shown as SEQ ID No.9, 7, 10 or a homologous sequence thereof, or are respectively shown as SEQ ID No.6, 7, 11 or a homologous sequence thereof.The antibody can specifically recognize a target antigen HpFucT protein, has high affinity, and can effectively inhibit the enzyme activity of the HpFucT protein.The antibody can be used for preparing a composition for detecting or diagnosing a disease related to the HpFucT protein, a product for purifying the HpFucT protein, a product for detecting the HpFucT protein level, a cell labeling product, a product for inhibiting the HpFucT enzyme activity, and a product for treating a disease related to the HpFucT protein.

The invention relates to the technical field of saccharomyces cerevisiae genetic engineering transformation, in particular to a high-throughput saccharomyces cerevisiae transformation method. The method comprises the following steps: screening an IGR sequence on a saccharomyces cerevisiae genome through bioinformatics, respectively taking 45bp from upstream and downstream of the IGR sequence as genome homologous sequences for primer design, designing and subpackaging a PCR premixed solution and the designed primer by using an automatic platform script for PCR amplification, adding an obtained PCR product into competent cells, adding a conversion solution, and carrying out PCR amplification on the competent cells to obtain the saccharomyces cerevisiae gene. Performing heat shock; washing and resuspending the cells subjected to heat shock, and then coating the cells on a screened solid culture medium for culturing; the conversion process is carried out on an automatic platform island. According to the method, the transformation efficiency is improved, the operation time of high-throughput transformation is shortened, the intelligence and stability of each transformation are improved, and the method is suitable for large-scale yeast genome editing, functional gene screening and saccharomyces cerevisiae genome site screening.

The present application relates to the technical field of bioengineering, and particularly relates to a kind of omega-transaminase mutant and its application.The present application discloses a kind of omega-transaminase mutant and its application in the synthesis of silodosin intermediate, the omega-transaminase mutant is by from Arthrobacter, through homologous modeling, reaction intermediate molecular docking, active site amino acid analysis, homologous sequence analysis, virtual mutation analysis combined with site-directed mutation verification, the single-point mutation of 62th and 223th obtains H62A, S223A mutant.The omega-transaminase mutant of the present application can one-pot catalysis 60mM silodosin intermediate ketone is converted into silodosin intermediate, conversion rate is greater than 92%, compared with wild type, the specific activity of H62A mutant to silodosin intermediate ketone is 1.4 times of wild type, S223A is 1.24 times of wild type, and the optimum reaction temperature is 30 DEG C, and the ee value of product is greater than 99.29%.

The present application relates to the technical field of drug target prediction, and discloses a drug target intelligent prediction method and system, the drug target intelligent prediction method comprising the following steps: step 1: obtaining structural data and sequence data of a target protein and homologous sequence data of the target protein; step 2: performing molecular dynamics simulation according to the structural data to calculate a dynamic conformation feature vector; calculating an evolutionary information feature vector according to the sequence data; obtaining an interaction feature vector by analyzing the structural data; step 3: obtaining an enhanced dynamic conformation feature vector, an enhanced evolutionary information feature vector and an enhanced interaction feature vector through an attention fusion mechanism, and then obtaining importance weights respectively through a gating fusion mechanism, and obtaining a fusion feature vector by element-by-element multiplication combined with the importance weights; step 4: inputting the fusion feature vector into a graph neural network prediction model to obtain a target druggability probability.

The application discloses a high-salinity wastewater anaerobic ammonia oxidation treatment method based on gene editing red bacteria, and belongs to the wastewater treatment and gene editing fields.The culture method of the anaerobic ammonia oxidation bacteria comprises the following steps: inoculating an original strain 16S rRNA into an anaerobic culture medium to culture; adopting a CRISPR-Cas9 gene editing technology, taking the nucleotide sequence described in SEQ ID NO:1 as a target sgRNA sequence, and precisely cutting and editing a target gene of the original strain; performing directional repair on the cut gene through a homologous sequence shown in SEQ ID NO:2; and culturing and purifying to obtain the target anaerobic ammonia oxidation bacteria.Through the CRISPR-Cas9 technology, the sgRNA is used as a guide molecule, the target gene is precisely cut and edited, the gene sequence is optimized through the homologous directional repair, and the ammonia oxidation capacity is enhanced; compared with the north discharge "red bacteria", the denitrification efficiency is increased by 4-6 times, and the cost is reduced by more than 60%.

The application discloses a synthetic protein design method based on protein feature engineering and diffusion model. First, a P-BLAST tool is used to search for homologous sequences of a query sequence in a non-redundant NR database by taking a target protein sequence as the query sequence; feature engineering is performed on the screened homologous sequences by using ProtParam-7 and ProtFactor-5; data represented by the ProtParam-7 and ProtFactor-5 features are taken as original protein input diffusion model; multidimensional screening is performed on the generated candidate proteins; wet experiments are adopted to evaluate the functional activity and structural stability of the candidate proteins, and proteins meeting the conditions are taken as the final generated results. The method combines protein sequence feature extraction and diffusion model generation, and through multidimensional screening and experimental verification, it is ensured that the generated protein is close to the target protein in structure and function.