75 results about "Prokaryote organisms" patented technology

The invention relates to a pTerm-SC plasmid as well as a construction method and application thereof. The pTerm-SC plasmid comprises a repE gene, a sopA gene, a sopB gene, a sopC gene, an ori2 replicon, an oriV replicon, prokaryote transcription terminators and antibiotics resistance genes. The pTerm-SC plasmid has the characteristics of very low copy number, high volume and stability and the like, can be applied to gene engineering fields such as the cloning, the screening, sequencing and the genome construction of complex-structure genes including repetitive-sequence genes, instable genes and long-fragment genes and has important role in the high-efficiency and high-quality synthesis of genes.

The invention belongs to the technical field of molecular biology, and discloses prokaryote-derived argonaute protein and application thereof. The prokaryote-derived argonaute protein is derived frommedium-temperature prokaryote kurthia masiliensis, and the prokaryote-derived argonaute protein is named as KmAgo. According to the protein and the application, polypeptide, nucleic acid, an expression vector, a composition, a kit and a method can perform site-specific modification on intracellular and extracellular genetic substances so that the protein can be effectively applied to many fields of biotechnology, such as nucleic acid detection, gene editing and gene modification, and a new tool is provided for gene editing, modification and molecular detection of the prokaryote-derived Argonaute polypeptide.

The invention discloses detoxication engineering bacteria, and a preparation method and application thereof. The invention provides a recombinant plasmid which is obtained by introducing coded genes of detoxication esterase and coded genes of hydrolase into an enzyme cutting site in plasmid pETDuet-1. The engineering bacteria of the invention can be obtained by introducing the recombinant plasmidinto host bacteria. The engineering bacteria comprise the coded genes of the detoxication esterase and the hydrolase and can realize inducible expression of the detoxication esterase and the hydrolase. Crude enzyme solution obtained by fermentation of the engineering bacteria can also be protected. The engineering bacteria and the crude enzyme solution thereof can simultaneously degrade organic acid esters such as malathion, parathion, high-efficiency cypermethrin, acetofenate and the like and organophosphorus pesticides, and can be used for detoxication of organophosphorus poisoned warm-blooded animals. The invention provides a new way for biological pollution treatment of toxic and harmful compounds such as pesticides remained in water sources, soil and the like by using natural resources of eukaryote and prokaryote.

The invention discloses an expression vector and a method for detecting membrane protein interaction in bacteria. The method comprises the following steps of firstly, constructing a double-label protein expression vector with a maltose binding protein label (MBP) and a histidine label (8*His), and expressing and purifying insoluble membrane protein by using the vector to increase the solubility ofthe membrane protein; and then, detecting the interaction between the membrane protein and other proteins by utilizing a pull down technology. Meanwhile, the method can also be used for screening proteins interacting with the membrane protein in bacterial whole-cell proteins. The method for detecting the interaction of the membrane proteins in the bacteria breaks through the technical difficultyof insolubility of the membrane proteins in prokaryotes, provides an experimental means for researching the functions of the bacterial membrane proteins, and has important application value.

The invention discloses an operon, a carrier thereof and application thereof. The sequence of the operon at least comprises any basic group of 201-257 bp, whose 3' ends are located in the upstream place of a corynebacterium glutamicum SEQ ID NO: 6 gene coding sequence ATG, and any basic group of 308-345 bp, whose 5' ends are located in the upstream place of the corynebacterium glutamicum SEQ ID NO: 6 gene coding sequence ATG. After the operon disclosed by the invention is induced by an inducing agent, the protein expression activity of the operon is obviously higher than the induced expressionactivity of a traditional Ptac; and the operon is an ultra-efficient operon. The operon disclosed by the invention can be used for efficient expression of procaryotic organism endogenous proteins orheterologous proteins.

The invention relates to recombination expression of cynoglossus semilaevis antibacterial peptide hepcidin and application thereof. A nucleic acid fragment used for recombining and expressing the cynoglossus semilaevis antibacterial peptide hepcidin with the sequence of SEQ ID NO: 1 is inserted into an expression vector, recombinant protein of the cynoglossus semilaevis antibacterial peptide hepcidin is expressed in host bacteria, a simple and effective method for purifying the recombinant protein is found out, and the protein expressed by the recombinant protein is applied to antibacterial products. In the invention, the vector and the host bacteria capable of carrying out antibacterial peptide recombination expression are constructed and sifted, the fusion expression and the simple and effective purification of the cynoglossus semilaevis antibacterial peptide in prokaryote are realized; compared with the traditional polypeptide synthesis method and the method for extracting polypeptide from natural resources, the preparation method provided by the invention makes large scale production in low cost possible and can be applied to feedstuff additives of marine cultivation fish, cultivation livestock and control of bacterial diseases.

The invention discloses a prokaryote-derived Mbp_Argonaute protein and application thereof. The Mbp_Argonaute protein has an amino acid sequence shown in SEQ ID NO: 1 or as shown in a sequence which has at least 50% or at least 80% of homology with SEQ ID NO: 1. According to the invention, an Argonaute protein gene derived from a cold-resistant prokaryotic organism Mucilaginibacter paaluis is synthesized, the protein is named as MbpAgo, and researches find that the MbpAgo has binding activity to single-stranded guide DNA and has nuclease activity to target RNA and/or target DNA complementarily paired with the single-stranded guide DNA, therefore, the MbpAgo can be used for targeted RNA editing in vivo and in vitro, and then specific site modification of genetic material. The MbpAgo not only can modify RNA with a high-grade structure, but also does not influence an endogenous RNAi pathway of animal and plant cells, provides a brand new powerful tool for RNA editing, and is high in cutting activity and good in specificity.

The invention proposes preparing biological samples for spectrometry which contain cell structures and/or whole cells of human or animal origin (e.g. thin human and animal tissue sections) or prokaryotes (e.g. microorganisms), and which require constant relative humidity, in a temperature-controlled gas volume whose humidity is determined by a saturated substance solution, for example a suitable salt solution. The invention exploits a physico-chemical phenomenon called “deliquescence”, which manifests itself by keeping the relative humidity above the saturated substance solution constant with a high degree of precision when a specified temperature is maintained. Pure succinic acid exhibits deliquescence at approx. 99% relative humidity, for example. Since an enormous variety of deliquescent salts and other suitable substances are available, it is possible to find the suitable substance for almost any desired relative humidity, with adjustment of the temperature, where necessary.

The present invention provides live recombinant microorganisms such as prokaryote organisms, particularly enterobacteria, preferably Salmonella enterica, containing SEQ. ID. no. 1 and SEQ. ID. no. 2 capable of expressing VapG and/or VapA/VapG lipoproteins (SEQ. ID. no. 3 and SEQ. ID. no. 4), optionally in combination with other active ingredients, methods for preparing vaccine strains, antigens (SEQ. ID. no. 3 and SEQ. ID. no. 4) and vaccine compositions, preferably vectored vaccines. Additionally, the present patent application relates to the use of the vaccine vectors in the preparation of pharmaceutical compositions, particularly vaccines indicated for the prevention and/or treatment of infections by Rhodococcus equi, its antibodies and/or antisera, diagnostic kits and methods for the prophylaxis and/or treatment of infections by R. equi.

The invention discloses a prokaryotic exosome and an extraction and separation method and application thereof, which belongs to the technical field of exosome extraction methods. The method comprisesthe steps of carrying out fermentation culture on prokaryotes, and centrifuging to acquire fermentation liquor; and carrying out multiple times of ultracentrifugation on the fermentation liquor to finally acquire the prokaryote exosome. The characteristics of the prokaryote exosome are identified through particle size analysis and nano transmission electron microscope analysis. According to the invention, the particle size of the prokaryotic exosome extracted and separated by the method integrally meets the standard, and the particle concentration can reach E + 9 or above; a clear vesicular structure can be seen in the field of view of an electron microscope, and the particle size of vesicles meets the detection standard of the exosome; the prokaryotic exosome extracted and separated by the method can be applied to transcriptomics, proteomics and lipidomics correlation analysis and preparation of an exosome drug loading system.

This invention provides a process for control in oil and gas wells and related facilities of prokaryote caused souring, fouling and corrosion by reduction of problematic prokaryotes with naturally occurring lysing organisms, particularly sulfate-reducing prokaryotes by proliferating suitable virulent lysing organisms under conditions in which problematic prokaryotes thrive, including in a gas production wellbore. The process provides in situ proliferation of virulent lysing organism in a wellbore by providing both virulent lysing organisms and their host prokaryotes to selectively grow an effective control amount and concentrations of lysing organisms in a well formation.

The invention relates to a low-copy pTerm plasmid and its construction method and application. The low-copy pTerm series plasmids include rop gene, rep gene, prokaryote transcription terminator and antibiotic resistance gene. A low-copy pTerm series plasmid of the present invention has the characteristics of low copy number, large capacity, and high stability, and can be used for cloning, screening, sequencing, and genome construction of genes with complex structures, including repetitive sequences, unstable genes, and long-segment genes In the field of genetic engineering, it plays an important role in the high-efficiency and high-quality synthesis of genes.

Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated proteins, which may be utilized in vaccines, including anti- bacterial vaccines. The glycosylated proteins may include a bacterial polysaccharide conjugated to a carrier, which may be utilized to generate an immune response in an immunized host against the polysaccharide conjugated to the carrier. Suitable carriers may include but are not limited to Haemophilus influenzae protein D (PD), Neisseria meningitidis porin protein (PorA), Corynebacterium diphtheriae toxin (CRM 197), Clostridium tetani toxin (TT), and Escherichia coli maltose binding protein, and variants thereof.

The invention discloses a film apparatus for culturing algae and prokaryotes, a bioreactor and a bioreactor system. The film apparatus comprises two opposite inner surfaces and the opposite inner surfaces comprise several separated seams, an individual cavity is formed by every two adjacent separated seams, the separated seams can not incompletely separate two cavities, bottoms of all individual cavities are communicated to form a first unimpeded connection cavity; and the bioreactor is composed of several film apparatus through arrangement and connection. The bioreactor system is composed of the bioreactors. The algae culture apparatus, the bioreactor and the bioreactor system have the advantages of low manufacture cost and no space restriction, and can conveniently and economically realize large scale microalgae culture.