30 results about "Gene assembly" patented technology

A viral vector production system is provided which system comprises:(i) a viral genome comprising at least one first nucleotide sequence encoding a gene product capable of binding to and effecting the cleavage, directly or indirectly, of a second nucleotide sequence, or transcription product thereof, encoding a viral polypeptide required for the assembly of viral particles;(ii) a third nucleotide sequence encoding said viral polypeptide required for the assembly of the viral genome into viral particles, which third nucleotide sequence has a different nucleotide sequence to the second nucleotide sequence such that said third nucleotide sequence, or transcription product thereof, is resistant to cleavage directed by said gene product. The viral vector production system may be used to produce viral particles for use in treating or preventing viral infection.

The invention relates to a gene assembly method of phage genetically engineered antibody library, belonging to the field of biomedical study and clinical applications. The method aims to overcome the problems of the prior art, including poor stability, low efficiency, high mutation probability and low fidelity, and indirect application to humanized single chain antibody study. The technical proposal adopted by the invention comprises the following steps of: obtaining heavy chain variable region gene and light chain variable region gene of a humanized antibody by RT-PCR, and ligating the antibody heavy chain variable region, a linker and the antibody light chain variable region with the pre-designed restriction enzyme cutting sites on two ends of primers using PCR and restriction enzyme method at the same time.

A recombinant operon comprising a gene assembly wherein there are at least two structural genes coding for at least two major subunits of colonization factor antigens (CFs) associated with enterotoxigenic Escherichia coli bacteria (ETEC), is disclosed. Further disclosed is a host cell, such as an Escherichia coli cell, genetically engineered to comprise such a recombinant operon, wherein said operon is located on an episomal element, such as a plasmid, or integrated in the chromosome of said host cell. Also disclosed is a method of producing a host cell capable of expressing from said operon at least two major subunits of colonization factor antigens (CFs) associated with enterotoxigenic Escherichia coli bacteria (ETEC). In addition, a vaccine composition against diarrhea comprising at least one such host cell together with pharmaceutically acceptable excipients, buffers, and/or diluents is disclosed. Finally is disclosed the use of said operon in the production of such a vaccine.

The present invention is directed to methods and compositions for use of homologous recombination for directed evolution, gene reassembly, and directed mutagenesis. One aspect of the present invention relates to methods for use of bacterial conjugative transfer and homologous recombination for directed evolution, gene reassembly, and directed mutagenesis. Another aspect of the present invention relates to compositions for use in or produced by the methods of the present invention, including libraries, archives and databases.

The invention discloses a recombinant CL7-CVN protein, and a preparation method and applications thereof. The preparation method includes cloning a coding sequence on an expression vector to obtain arecombinant expression vector; and converting the recombinant expression vector into host cells to perform expression and purification so as to obtain the recombinant CL7-CVN protein. Through a synthesized CVN gene and gene assembly method, the fusion expression vector can be built, and the highly soluble expression of the vector in escherichia coli can be realized; and according to the heat resistance of the recombinant CL7-CVN protein and through heat treatment and 3C protease digestion, the recombinant CL7-CVN protein and CVN protein can be rapidly obtained through one-step affinity chromatography, and purity can reach 99%. The provided preparation method can make the recombinant protein expression significantly enhanced, make activity more stable and make a purification method simpler;and both the prepared recombinant CL7-CVN protein and CVN protein have antiviral activity.

The invention discloses a novel method for gene assembly using improved CRISPR-Cpf1. The method includes utilizing a DNA gene editing enzyme FnCpf1 from a Francisella tularemia noticed U112 strain (gene sequence number: MF193599) for gene recombinant cloning. The seed sequence of crRNA (an RNA sequence generated by transcription of regular clustering spaced CRISPR) from 23 nucleotides reported inthe literature to 18 without affecting the FnCpf1 cleavage efficiency. At the same time, oligo(dN)6 (oligonucleotide hexamer) is used as the viscous end of the cleavage target site, the problem of terminal heterogeneity after FnCpf1 digestion is solved, the assembly efficiency of exogenous DNA fragments is remarkably improved, and the DNA fragment assembly efficiency reaches about 85%. The gene editing enzyme FnCpf1 can be replaced with other nucleic acid editing enzymes Cpf1 such as AsCpf1 and LbCpf1.

The invention relates to an assembling method for de novo sequencing data based on an optics map platform Irys. The method comprises: using the optics map platform Irys to obtain a gene assembly file; getting a scaffold file: fai file of NGS; preprocessing data: through setting a threshold value, filtering a comparison result whose confidence level is low, combining cmap files, sorting, calculating N50; counting assembling effects: counting comparison results of BioNano and NGS, including the contig of the BioNano, and the length, number, and total quantity of scaffold of NGS; according to the contig of the BioNano and network topological relations among the scaffold of the NGS, analyzing length of the assembled new contig and length of scaffold in a classified manner. The method can assist genome assembly, and obviously improves gene assembly effects of species.

A multi-gene carrier system is composed of a receptor carrier and at least two donor carriers, and is prepared by altermative gene assemblings with the said carriers. It can be used for conversion of multiple genes to obtain different genetically engineered products and multi-gene expression characteristics.

The present invention provides an error correcting method of test sequence, which involves receiving test sequences, configuring high frequency short string list based on a preset high frequency threshold value, traversing each received test sequence, searching an area with the largest number of continuous high frequency short strings on each test sequence in combination with high frequency short string list, configuring whole left sequence and/or right sequence of high frequency short strings at left side and/or right side of searched area according to corresponding received test sequence and high frequency short string list, and constituting corresponding test sequence according to configured left and/or right sequence and searched area. The present invention also provides corresponding error correcting system of test sequence and gene assembly equipment.

The invention discloses a method for improving caffeic acid content and/or biological self-luminous intensity of plants, and belongs to the technical field of biology. The method comprises the following steps: (1) sequentially integrating an NtCM1 gene, an NtPAT gene, an NtADT2 gene, an NtPAL1 gene and an NtC4H gene into a receptor vector by utilizing a multi-gene assembly technology, and constructing to obtain a multi-gene vector; and (2) introducing the target gene segment into a receptor plant by using a transgenic technology to obtain a transgenic plant with enhanced caffeic acid content and/or biological self-luminous intensity. The gene participates in synthesis of caffeic acid in the plant for the first time, and co-expression of the gene and a self-luminous Hisps gene, a CPH gene, an H3H gene, an NPGA gene and a Luz gene is carried out, so that the biological self-luminous intensity of the plant can be remarkably improved. The invention provides a feasible technical scheme for breeding and production of high caffeic acid plants and sustainable luminous plants.

The present invention is directed to methods and materials for RNA-mediated gene assembly from oligonucleotide sequences. In some embodiments, the oligonucleotides used for gene assembly are provided in an array format. An RNA polymerase promoter is appended to surface-bound oligonucleotides and a plurality of RNA copies of each oligonucleotide are then produced with an RNA polymerase. These RNA molecules self-assemble into a desired full-length RNA transcript by hybridization and ligation. The resulting RNA transcript may then be converted into double strand DNA useful in a variety of applications including protein expression.

The invention provides an informatics-based gene transcript assembly and quantification method and system, wherein the method comprises: aligning a sequencing read with a reference genome, and predicting initial start and end positions of a gene and transcript according to the alignment results for the sequencing read and reference genome; after prediction, establishing a directed graph to simulate possible transcripts to obtain a candidate transcript set; subjecting the candidate transcript set to transcript prediction and kurtosis estimation according the mode of maxium information transmission capacity. The method and system of the invention have the advantages that dependence on external gene positional marking is avoided, gene assembly accuracy is significantly improved, and sequencing precision is improved.

A construction method of a stem cell bank for preventing and treating novel coronavirus pneumonia comprises the following steps of: collecting residual amniotic fluid cells, newborn umbilical cord blood, umbilical cord and placenta tissues after prenatal diagnosis, carrying out stem cell induction, immortalized gene transfection, novel coronavirus targeted interference gene assembly, novel coronavirus susceptibility gene knockout or knock-in and novel coronavirus antibody generation gene assembly to prepare immortalized stem cells, DNA recombinant stem cells, RNA interference stem cells, gene silencing stem cells and various stem cell vector vaccines for preventing and treating novel coronavirus pneumonia, collecting the prepared stem cell products in -196 DEG C liquid nitrogen in a centralized mode according to donor names, ABO blood types or HLA types, and establishing a renewable special stem cell bank. When the novel coronavirus is epidemic, various types of stem cells or vaccines thereof can be extracted from the stem cell bank to be cultured and amplified, and individualized treatment is performed by the patient's own or homologous stem cells, so that side effects such as immunological rejection and the like of traditional stem cell or vaccine prevention and treatment are reduced, and the problem of lack of COVID-19 special-effect medicines and stem cells is solved.