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6 results about "Cloning vector" patented technology
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A cloning vector is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. The vector therefore contains features that allow for the convenient insertion or removal of a DNA fragment to or from the vector, for example by treating the vector and the foreign DNA with a restriction enzyme that cuts the DNA. DNA fragments thus generated contain either blunt ends or overhangs known as sticky ends, and vector DNA and foreign DNA with compatible ends can then be joined together by molecular ligation. After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.
Assembling synthetic DNA constructs from natural DNA
The disclosure provides for methods of constructing synthetic chromosomes including the steps of providing host cells with an endogenous chromosome, transforming a cloning vector and a cloning cassette into the host cells, excising target genomic nucleic acids from the endogenous chromosome, recombining the excised target genomic nucleic acids with the cloning cassette via homologous recombination to form heterologous vectors comprising cloned sequences, extracting the heterologous vectors containing the cloned sequences from the host cells, digesting the heterologous vectors with a restriction endonuclease to release the cloned sequences from the heterologous vectors to provide released cloned sequences, and introducing the released cloned sequences, a centromere cassette, and a yeast artificial chromosomes or bacterial artificial chromosomes into a second host cell, such that the released cloned sequences, the centromere cassette, and the YAC or BAC recombine with one another via homologous recombination to produce the synthetic chromosome.
Owner:UNIV OF SOUTHERN CALIFORNIA
Determination method of multivalent antigen sequence of anti-periodontitis red complex
PendingCN122081364AImproving immunogenicityHighly conservativeBacterial antigen ingredientsHydrolasesEnzyme digestionTGE VACCINE
The invention belongs to the technical field of biology, and discloses a method for determining a multivalent antigen sequence of an anti-periodontitis red complex, which comprises the following steps: firstly, screening out key virulence factor antigens (FimA-II, RgpA, Msp and BspA) from three core pathogenic bacteria of the red complex: porphyromonas gingivalis, treponema denticola and fusisterone; then, independently cloning each antigen gene to a pUC57 cloning vector, constructing a single antigen recombinant plasmid, and avoiding intergene interference by designing a specific enzyme cutting site; and finally, through plasmid transformation, positive clone screening and double enzyme digestion verification, the plasmid quality is ensured. The obtained single antigen plasmid can be flexibly combined and used, is beneficial to subsequent multivalent vaccine development, and has the advantages of wide antigen coverage, high expression reliability, flexible construction strategy, easiness in industrialization and the like.
Owner:JINYUE ZHICHENG (LIAONING) BIOTECHNOLOGY CO LTD
Smhrd3a gene and application of the same in increasing tanshinone content in salvia miltiorrhiza
PendingCN122081339AIncrease tanshinone contentFermentationVector-based foreign material introductionBiotechnologySalvia miltiorrhiza
This invention belongs to the field of genetic engineering technology and provides the SmHRD3A gene and its application in increasing the tanshinone content in *Salvia miltiorrhiza*. This invention comprehensively applies transcriptome data analysis, gene cloning, vector construction, molecular detection, RT-qPCR analysis, target exploration, protein-protein interaction, and compound content determination to obtain the encoding gene of the ERAD protein SmHRD3A, which interacts with the rate-limiting enzyme SmHMGR in the MVA pathway. This gene can significantly increase the tanshinone content in *Salvia miltiorrhiza*. It provides new ideas and target genes for the production of active ingredients in *Salvia miltiorrhiza* and for molecular design breeding of *Salvia miltiorrhiza*, and has high application and research value.
Owner:THE NAVAL MEDICAL UNIV OF PLA
Application of slgrp gene and encoded protein in regulating plant resistance to tomato brown rugose fruit virus and transgenic plant breeding method
The application discloses a kind of SlGRP gene and the application of coding protein in regulating plant resistance tomato brown rugose fruit virus and transgenic plant cultivation method, wherein the transcript sequence of SlGRP gene is as shown in SEQ ID NO:1.The application constructs the cloning vector of pCAMBIA1300-SlGRP-GFP using homologous recombination system, amplifies gene SlGRP using the cDNA of tomato, connects SlGRP into pCAMBIA1300-GFP vector by homologous recombination, so as to obtain the overexpression vector containing SlGRP, imports gene into tomato, tobacco and other plants by the method of agrobacterium transformation and obtains stable inheritance overexpression strain, and through disease resistance determination, it is shown that the overexpression plant of SlGRP can significantly inhibit the infection of tomato brown rugose fruit virus.
Owner:INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
Method for constructing target DNA sequences and cloning vectors
The present application relates to a method for producing a target DNA sequence and a cloning vector, comprising the steps of amplifying and extracting a DNA construct in a host cell and a protelomerase-type IIS restriction endonuclease and / or meganuclease-DNA exonuclease catalyzed three-step isothermal enzymatic reaction step, where the construct is autonomously replicating and contains (a) one or more type IIS restriction endonuclease and / or meganuclease recognition sequences; (b) a target DNA sequence; and (c) protelomerase recognition sequences at the two terminal flanks of the target DNA sequence.
Owner:NANJING GENSCRIPT BIOTECH CO LTD