11 results about "Restriction site" patented technology

This invention belongs to the field of molecular biology detection, specifically relating to a rapid detection method for economic traits of Tibetan chickens. FBXO32 Gene SNP markers, detection methods, and applications. This invention is the first to discover: Tibetan chickens... FBXO32 The G / A mutation at position 8916 of the gene is closely related to the presence or absence of the Hpa II restriction site, and is also closely related to the main economic traits of Tibetan chickens. Based on this new discovery and the SNP site, this invention provides a method for rapidly detecting this SNP site and screening for economic traits in Tibetan chickens. The method is simple, convenient, fast, low-cost, and provides reliable results.

The invention relates to a Pseudomonas sp. strain for use in the production of a recombinant protein characterised in that said strain comprises a nucleotide sequence encoding a phi15 RNA polymerase. The invention further relates to a plasmid, capable of integrating or replicating in Pseudomonas sp., comprising a phi15 promoter sequence operably linked to a nucleotide comprising one or more restriction sites for the insertion of a nucleotide sequence encoding a recombinant protein, or operably linked to a nucleotide sequence encoding a recombinant protein.

The invention discloses a urate oxidase gene derived from aspergillus cristatus and application thereof, and the urate oxidase gene is characterized in that a natural urate oxidase gene coding region sequence is obtained from the aspergillus cristatus, and the gene is optimized according to the codon preference of escherichia coli to obtain an optimized sequence. Connecting the optimized gene to a carrier through NdeI and BamHI restriction enzyme cutting sites, and constructing a wild type uox recombinant plasmid; the method comprises the following steps: replacing a sequence of uox of aspergillus cristatus for coding 243-site tyrosine (Y) with a sequence for coding lysine (K) through site-specific mutagenesis, constructing mutant Y243K recombinant plasmids, respectively transforming the recombinant plasmids into competent cells, culturing, and adding IPTG to induce target protein expression. And collecting the induced thalli, carrying out ultrasonication and centrifugation to collect the supernatant so as to obtain a crude enzyme, and purifying the crude enzyme through a nickel affinity column so as to obtain the purified UOX protein. Uric acid is used as a substrate to measure the enzyme activity of the purified UOX, and the result shows that the specific enzyme activity of the mutant Y243K is obviously higher than that of a wild type, and the activity is improved by 2.75 times, so that the mutant Y243K has higher application value.

This invention provides a research model and construction method for calcium ion imaging of ASH neurons in nematodes, belonging to the field of biological model construction technology. The construction method includes the following steps: nematode culture; construction of a transgenic probe plasmid: obtaining the ASH neuron-specific promoter sra-6 using PCR technology, and connecting HindIII and BamHI restriction sites to both ends of the promoter; amplifying YC3.60 using PCR technology; then inserting the promoter sequence into the pPD95.75 plasmid, and then inserting YC3.60 after the promoter. Transgenic microinjection of nematodes is used, followed by screening to obtain nematodes with ASH neurons carrying green fluorescent protein. In this application, YC3.60 is transferred into nematodes via transgenic means. Using a promoter specifically expressed in ASH neurons to link the gene of this fluorescent probe protein into the nematode, the fluorescent probe can be specifically expressed in the neuron. The constructed nematode model can be used for neuronal structure and functional imaging studies, as well as neuropharmacological efficacy analysis.

The invention relates to the technical field of viruses, in particular to a construction method and application of a porcine parvovirus full-length infectious clone carrying a stable genetic marker. The method comprises the following steps: cloning a complete genome of a porcine parvovirus SXND18 strain to a plasmid pBluescript II SK (+) through an In-fusion seamless cloning technology, so as to obtain a recombinant plasmid pSXND18; meanwhile, a new EcoRI restriction enzyme cutting site is introduced into a genome by utilizing an overlapping PCR (Polymerase Chain Reaction) technology and is used as a genetic marker for identifying a wild virus and a rescue virus; and mixing the recombinant plasmid with a transfection reagent, and transfecting ST cells to obtain the rescue virus rSXND18. The porcine parvovirus reverse genetic operating system established by the invention can be used for research in the directions of porcine parvovirus virulence analysis, cross-species transmission mechanism and the like, and also lays a foundation for research on novel vaccines of the porcine parvovirus.

The invention provides construction and application of a medical intermediate P29 eukaryotic expression engineering bacterium, and belongs to the technical field of biosynthesis, the engineering bacterium takes pichia pastoris as a chassis cell and contains a recombinant plasmid for expressing a fusion protein, and a gene sequence of the fusion protein contains a protection sequence-enzyme cutting site-P29 sequence structure. Wherein the protection sequence is derived from collagen and is specifically shown as SEQ ID NO. 2. According to the invention, a medical intermediate P29 eukaryotic expression engineering bacterium is successfully constructed by using pichia pastoris, and the engineering bacterium can realize efficient and low-cost production of the intermediate P29, and provides a substrate and a raw material for chemical synthesis of the semeglutide.

A recombinant polynucleotide suitable for use in a display vector is provided. The recombinant polynucleotide includes from 5′ to 3′: a first nucleic acid sequence (or insert) encoding an amino acid sequence to be displayed on a surface; a first pre-selected restriction site; a second nucleic acid sequence encoding a surface peptide capable of being displayed on the surface; and a second pre-selected restriction site. Corresponding display vectors that can be converted into expression vectors in a high-throughput fashion, as well as methods of use thereof, are also provided.

The invention provides a fusion protein which is characterized in that the fusion protein comprises the following elements from the 5'end to the 3 'end: 5'-MBP-Furin restriction enzyme cutting site-rhIL-7-3 ', and a method for in-vitro expression and purification of recombinant human interleukin 7 in mammalian cells, and the method comprises the following steps: a, providing a first expression vector, the first expression vector comprises nucleic acid for coding the fusion protein; b, providing a second expression vector, wherein the second expression vector comprises nucleic acid for coding furin; c, co-transfecting the first expression vector and the second expression vector to mammalian cells, and performing fermentation culture to enable the cells to express recombinant human IL-7 protein; d, centrifuging to obtain fermentation supernate from the step c; and e, purifying by using POROS XS and CHT chromatographic methods to obtain the rhIL-7 fragment.

The invention relates to the technical field of plant genetic transformation, and provides a plant genetic transformation system based on direct delivery, a genetic transformation method and application. An original vector pMKV057 is directionally modified to obtain a modified vector, and the modification method comprises the following steps: replacing an LUC expression cassette of the vector with a modular expression cassette integrated with an epitope tag; the method comprises the following steps: designing multiple cloning sites on a vector, introducing restriction enzyme cutting sites HindIII and EcoRI or HindIII and XbaI, and inserting a Kozak sequence into a 5'end upstream adjacent region of a nucleotide sequence of the multiple cloning sites to obtain a modified vector. And transforming the modified vector into agrobacterium tumefaciens to carry out plant genetic transformation. Through the technical scheme, the problems of low plant genetic transformation efficiency and long transformation time in related technologies are solved.

The present application relates to a method for analyzing the similarity of at least two samples of a plurality of samples comprising genomic DNA. The method comprises the following steps: a) providing a plurality of samples comprising genomic DNA; b) performing a deterministic restriction site whole genome amplification (DRS-WGA) of the genomic DNA separately for each sample; c) preparing a massively parallel sequencing library from each product of DRS-WGA using a no-fragmentation, sequencing adapter / WGA fusion primer PCR reaction; d) performing low-pass whole genome sequencing of the massively parallel sequencing library at an average coverage depth of less than 1x; e) aligning the reads of each sample obtained in step d) to a reference genome; f) extracting the allele content at a plurality of polymorphic loci for each sample; g) calculating a pairwise similarity score locus for at least two samples based on the measured allele content at the plurality of loci; h) determining the similarity of at least two samples based on the similarity score, the method being used for non-invasive prenatal testing or diagnosis.

This invention discloses a PTPσ_Ig1-3 fusion protein, its preparation method, and its applications, belonging to the field of genetic engineering. The recombinant fusion protein is obtained by adding an inducible peptide to the N-terminus of the PTPσ_Ig1-3 domain, and sequentially adding a TEV restriction site, GFP fluorescent protein, and a His tag to the C-terminus. The amino acid sequence of the PTPσ_Ig1-3 domain is shown in SEQ ID NO.1, the amino acid sequence of the inducible peptide is shown in SEQ ID NO.2, and the amino acid sequences of the TEV restriction site, GFP fluorescent protein, and His tag are shown in SEQ ID NO.3. Experimental verification showed that this recombinant fusion protein has significant binding activity with glycosaminoglycans such as heparin sodium and chondroitin sulfate, showing promising applications in the study of axonal injury and regeneration mechanisms and in drug delivery therapy.