305 results about "Synthetic biology" patented technology

The present invention provides oligonucleotide constructs, sets of such oligonucleotide constructs, and methods of using such oligonucleotide constructs to provide validated sequences or sets of validated sequences corresponding to desired ROIs. Such validated ROIs and constructs containing these have a wide variety of uses, including in synthetic biology, quantitative nucleic acid analysis, polymorphism and/or mutation screening, and the like.

This invention provides methods and compositions for assembling biological constructs (e.g., plasmids, transformed cells, etc.). In certain embodiments the methods involve encapsulating separate components of said biological construct each in a fluid droplet confined in a fluid channel; optionally mixing droplets from different fluid channels to for a sequenced order of droplets carrying different components of said biological construct in a channel or chamber; and optionally combining two or more droplets each containing different components of said biological construct to permit said components to react with each other in one or more reactions contributing to the assembly of said biological construct.

The invention discloses a bacterial strain for producing farnesene and an application of the bacterial strain, belonging to the field of synthetic biology. The bacterial strain for producing the farnesene contains related genes for synthesizing the farnesene through a mevalonic acid pathway and codon optimization; and the sequence of a farnesene synthetic gene afs optimized by codons is as shown by SEQ ID NO.1. The bacterial strain can be used for producing the farnesene, and a seed solution of the bacterial strain is inoculated into a culture medium containing a carbon source to carry out prokaryotic expression to obtain the farnesene. The gene for synthesizing the farnesene is subjected to codon optimization or the farnesene is synthesized by using the mevalonic acid pathway to ensure that each protein is closer to a ratio of AtoB:ERG13:tHMG1:ERG12:ERG8:MVD1:Idi:IspA:AFS=1:10:2:5:5:2:5:2:2, so that the production of the farnesene can be further promoted. By adopting the bacterial strain, the output of the farnesene is greatly improved to be more than 1g/L.

The invention provides a droplet encapsulate comprising: a drop of a hydrophobic medium; a peripheral layer of non-polymeric amphipathic molecules around the surface of the drop; and an aqueous droplet within the peripheral layer, the aqueous droplet comprising: (a) an aqueous medium and (b) an outer layer of non-polymeric amphipathic molecules around the surface of the aqueous medium. The invention also provides processes for preparing the droplet encapsulates. Various uses of the droplet encapsulates are also described, including their use as drug delivery vehicles, in synthetic biology, and in the study of membrane proteins.

The present disclosure generally relates to genetic engineering of bacteria. More particularly, the present disclosure relates to genetic engineering of Gram-negative bacteria expressing different species of lipid A on their surface. In one embodiment, the present disclosure provides for an engineered strain of E. coli according to Table 1. In another embodiment, the present disclosure provides for a lipopolysaccharide purified from an engineered strain of E. coli according to Table 1.

The invention discloses a phage display antibody library and five strains of screened antibodies capable of being combined with S protein of novel coronavirus SARS-CoV-2. Mutation is introduced into an ultra-variable region of an antibody variable region based on synthetic biology and a phage display technology, and a gene is transferred into escherichia coli, so that a synthetic antibody librarycontaining 108 kinds of antibodies is constructed; the phage display antibody library provided by the invention can perform screening to obtain the antibody with the specificity and the detection function, so that powerful resources of biological research and medical diagnosis are expanded; and the five strains of antibodies capable of being combined with the S protein of the novel coronavirus arefurther screened out and can be used for detecting the virus, part of the antibodies can block combination of the virus and cells, and the antibodies have the capacity of neutralizing novel coronavirus infectivity, can be used for preparing a novel coronavirus detection product, preparing a drug for inhibiting the novel coronavirus and preparing a pharmaceutical preparation for preventing or treating diseases caused by the novel coronavirus, and have a wide application prospect.

The invention relates to application of lysinibacillus boronitolerans in inhibiting growth of magnaporthe oryzea. The lysinibacillus boronitolerans not only can reduce the occurrence and hazard of therice blast to a great extent, but also has no influence on the ecological environment, is expected to serve as a high-quality chassis microorganism for synthetic biology researches, and brings a newdirection for control over the rice blast.

The invention relates to the field of genetic engineering tools, methods and techniques for gene or genome editing. Specifically, the invention concerns isolated polypeptides having nuclease activity, host cells and expression vectors comprising nucleic acids encoding said polypeptides as well as methods of cleaving and editing target nucleic acids in a sequence-specific manner. The polypeptides, nucleic acids, expression vectors, host cells and methods of the present invention have application in many fields of biotechnology, including, for example, synthetic biology and gene therapy.

The present invention relates generally to the fields of genomics, synthetic biology and genetic engineering. More particularly, the present invention concerns the methods that enable parallel multiplex ligation and amplification on surface for making assemblies of nucleic acids of various biological applications and for analysis of biological samples such as DNA, RNA, and proteins.

The invention discloses applications of glycosyltransferase and related materials thereof in the construction of engineering bacteria for producing ginsenoside Rb1 and Rg1. A glycosyltransferase genePn3-32 which can catalyze ginsenoside Rd to generate ginsenoside Rb1 can be successfully identified through a synthetic biological method; and the gene can simultaneously catalyze ginsenoside F1 to generate ginsenoside Rg1 and construct recombinant yeast producing the ginsenoside Rb1 and the ginsenoside Rg1. Through experiments, the constructed recombinant yeast producing the ginsenoside Rb1 and the ginsenoside Rg1 can simultaneously generate the ginsenoside Rb1 and the ginsenoside Rg1. Pn1-31, Pn-3-29, Pn3-31 and Pn3-32 glycosyltransferase genes in medicinal plant radix notoginseng are firstly utilized to continuously catalyze protopanaxadiol and protopanaxatriol to synthetize the ginsenoside Rb1, the ginsenoside Rg1 and corresponding intermediates, so that novel cases can be provided formicrobial strains to produce natural products.

The invention provides a hydrogel network comprising a plurality of hydrogel objects, wherein each of said hydrogel objects comprises: a hydrogel body, and an outer layer of amphipathic molecules, on at least part of the surface of the hydrogel body, wherein each of said hydrogel objects contacts another of said hydrogel objects to form an interface between the contacting hydrogel objects. A process for producing the hydrogel networks is also provided. The invention also provides an electrochemical circuit and hydrogel component for mechanical devices comprising a hydrogel network. Various uses of the hydrogel network are also described, including their use in synthetic biology and as components in electrochemical circuits and mechanical devices.

The invention discloses a genetic engineering stain taking tyrosine as a substrate to synthesize naringenin and a construction method thereof, belonging to the field of synthetic biology or metabolic engineering. According to the construction method disclosed by the invention, a synthesis route constituted by six genes, namely a gene matB encoding a malonic acid transporting enzyme, a gene matC encoding a malonic acid absorption route, a gene encoding a tyrosine ammonialyase (TAL), the gene encoding a 4-cinnamic acid: coenzyme A ligase (4CL), the gene encoding a chalcone synthase (CHS) and the gene encoding a chalcone isomerase (CHI) is introduced into Escherichia coli BL21 to obtain a recombinant strain capable of taking the tyrosine as the substrate to synthesize the naringenin, the synthesis route is further optimized through a modular transformation theory, the strain is finally fermented in a shaking flask for 72 hours by taking the tyrosine as the substrate, and the yield of the naringenin achieves 90mg/L. A strategy adopted by the construction method disclosed by the invention provides certain reference significance for future production of natural small molecular substances by a microbiological method.

The invention relates to a gene cluster participating in synthesis of cucurbitacin E of watermelon and an application of the gene cluster. According to the invention, the gene cluster participating in synthesis of cucurbitacin E is found in the watermelon genome for the first time, the gene cluster comprises eight genes, wherein six of the eight genes are positioned within the range of the No.6 chromosome 57kb. A yeast system is adopted for analyzing the reaction from the step 1 to the step 4 of the synthesis of cucurbitacin E. The invention further discloses a molecular mechanism for forming the bitter taste of the watermelon, so that a theoretical basis and a molecule assistant breeding objective are provided for the breeding of the watermelon without the bitter taste, and valuable experiences are provided for heterologous biosynthesis and development and utilization of cucurbitacine. The technology provided by the invention can replace the traditional method for extracting bitter principle from a plant material, so that synthesis of the bitter principle in vitro is realized by utilizing synthetic biology.

The present invention relates to a carotenoid cleavage dioxygenase CCD4a involved in the synthesis of gardenia jasminoides ellis crocin, an encoding gene and functions thereof, belongs to the technical field of gene engineering, and discloses the open reading frame sequence of a carotenoid cleavage dioxygenase CCD4a gene and the amino acid sequence encoded by the carotenoid cleavage dioxygenase CCD4a gene. According to the present invention, the CCD4a gene is screened based on transcriptomics, the functions of CCD4a are verified in vitro by using zeaxanthin as the substrate and constructing the pET28a-CCD4a prokaryotic expression vector, and the results prove that CCD4a has the functions of carotenoid cleavage dioxygenase so as to establish the foundation for the analysis and biosynthetic research of the crocin source pathway.

The invention discloses a pseudomonas putida gene knockout and genome simplification system, and belongs to the field of microbial gene engineering. According to the pseudomonas putida gene knockout and genome simplification system disclosed by the invention, a suicide plasmid knockout system and a site specific recombination system are skillfully combined, so that the correction rate of two turns of screening is greatly improved to be higher than 90%, and the correction rate is still higher than 90% when a gene cluster, which is 69 genes long, is knocked out. The system disclosed by the invention achieves the continuous knockout of the gene cluster, simple and convenient to operate, efficient, high in correction rate and low in cost. The system disclosed by the invention lays a molecular basis for the simplification and the optimization of a pseudomonas putida genome; therefore, the system is conducive to the implementation of efficient construction of chassis cells of synthetic biology.

The invention discloses a method for constructing a high-yield sclareol engineering strain by a synthetic biology methdo, and an application thereof. The construction method for the strain relates to express plant sourced key enzymes for synthesizing sclareol, wherein the key enzymes comprise a pyrophosphoric acid ladanum enediol ester synthase and a sclareol synthase; and overexpress a combination of one or more yeast endogenous enzyme-based genes, wherein the enzyme-based genes comprise a key enzyme of a mevalonic acid approach-(hydroxymethyl) coenzyme A reductase, and two enzymes of an isopentenylpyrophosphate approach-a farnesyl pyrophosphate synthase and a geranylgeranyl diphosphate synthase. The method can efficiently produce sclareol, with a shake flask fermentation level reaches 9 mg/l, and has industrial application prospects.

The invention belongs to the technical field of genetic engineering, and particularly relates to a multi-gene locus simultaneous editing method based on an endogenous CRISPR-Cas system of zymomonas mobilis and application thereof. The method mainly comprises the following steps of: constructing a plasmid containing an artificial CRISPR expression unit; selecting a plurality of target editing target loci, and designing a gRNA sequence; connecting a plurality of gRNA sequences in series, and constructing the gRNA sequences on the plasmid containing the artificial CRISPR expression unit; constructing a donor DNA sequence on a vector to obtain an edited plasmid; and transforming the edited plasmid into competent cells for editing. The method constructs a gene editing platform by utilizing theendogenous CRISPR-Cas system of zymomonas mobilis, breaks the limitation of low efficiency of exogenous CRISPR-Cas9 genome editing in the strain, realizes rapid and efficient knockout of multiple genes in the strain, and promotes the development of metabolic engineering, systemic biology and synthetic biology.

The invention discloses a terpenoid synthase for producing nerolidol and application thereof, and belongs to the field of synthetic biology. Terpene synthase TPF09930 for producing the nerolidol is provided, a nucleotide sequence of the terpene synthase TPF09930 is as shown in SEQIDNO:2, the terpene synthase TPF09930 is combined with mevalonic acid pathway related genes to construct a strain for producing the nerolidol. By overexpression of Escherichia coli XL1 blue-derived atoB gene or idi gene of mevalonic acid pathway, catalytic substrate Farnesyl pyrophosphate can be synthesized in large scale, and producing of the nerolidol can be further promoted. The terpene synthase has specificity and high efficiency, can improve the nerolidol yield, greatly overcomes the defects of a large amount of raw materials and a low nerolidol yield, reduces study cost, and ensures greenness and environmental friendliness.

The invention discloses a recombinant bacillus subtilis of high-yield pullulanase and a construction method thereof. The construction method of the recombinant bacillus subtilis of the high-yield comprises the following steps that an artificial operon BPB used for expressing the pullulanase is used for constructing a recombinant plasmid pGE-BPB; the nucleotide sequence of the artificial operon BPB is shown in the graph SEQ ID NO.4; the constructed recombinant plasmid pGE-BPB is converted into a bacillus subtilis competent cell, and through secondary recombination, a neutral protease gene nprE in the bacillus subtilis competent cell is replaced by the artificial operon BPB in situ. According to the recombinant bacillus subtilis of the high-yield pullulanase and the construction method thereof, firstly, the acidproof and heatproof pullulanase gene of an original bacterial strain is optimized, based on the synthetic biology method, a plurality of molecular elements capable of improving the gene transcriptional level are assembled into the artificial operon, and the recombinant bacillus subtilis can be obtained through construction. The recombinant bacterial strain can ferment to generate the high-yield pullulanase, and the enzyme activity each unit can reach or exceed 300 U/ml.

The invention belongs to the technical field of genetic engineering, particularly relates to a genome editing method based on a zymomonas mobilis endogenous CRISPR-Cas system and application thereof and aims at taking Z.mobilis 4 as a type strain and utilizing the I-F type CRISPR-Cas system coded by its genome to establish a genome editing platform. A powerful tool is provided for basic research and application research in the strain and similar cells, and the development of metabolic engineering, system biology and synthetic biology is promoted. According to the technical scheme, the genome editing method is characterized by comprising the steps of constructing a plasmid containing an artificial CRISPR expression unit; selecting a guide RNA sequence aiming at a target site; constructing aguide RNA primer sequence on the plasmid containing the artificial CRISPR expression unit; constructing a donor DNA sequence on a vector to obtain an editing plasmid; transforming the editing plasmidinto competent cells for editing.

The invention belongs to the fields of genetic engineering and synthetic biology, and in particular relates to general type CART/TCRT cells with antibody drug resistance as well as a construction method of the general type CART/TCRT cells. The general type CART/TCRT cells are allogeneic T cells having chimeric antigen receptors and T cell receptors, and alpha and beta chains and CD52 molecules ofthe T cell receptors are knocked out; a technology for knocking out the alpha and beta chains and the CD52 molecules of the T cell receptors is a CRISPR technology; the general type CART/TCRT cells not only retain the targeting of tumor specific antigen, but also eliminate the problems of GvHD and rejection, and weaken the sensitivity to an antibody drug Alemtuzumab; the general type CART/TCRT cells can be used as a general type simple, low-cost and high-activity CART/TCRT cell preparation.

The invention discloses a system and microorganism for producing taxadiene, and application of the system or the microorganism, and belongs to the field of synthetic biology. The system for producing the taxadiene comprises mevalonate pathway related genes or functional equivalents thereof, and taxadiene synthetized related genes or functional equivalents thereof. The mevalonate pathway related genes are selected from atoB and idi of escherichia coli BL21, and erg13, thmg1, erg12, erg8, and mvd1 of brewer's yeast INVSC1; gene nucleotide sequences of the taxadiene synthetized related genes ggpps are shown by SEQID No.1 and SEQID No.2. The microorganism for producing the taxadiene is the microorganism containing the system for producing the taxadiene, by applying the system into the microorganism, the microorganism for producing the taxadiene is obtained. The microorganism produced in the invention can produce culture for the taxadiene stably, lays a foundation for further using the microorganism to produce paclitaxel, and has a wide application prospect.

A novel synthetic biology-based ADCC technology is provided that enhances or enables ADCC response. The novel ADCC technology can be used to prevent or treat cancers, infectious, inflammatory or autoimmune diseases, and other diseases where elimination of diseased cells is desirable.