146 results about "Bacterium coli" patented technology

The invention provides nucleic acid sequences which are complementary, in one embodiment, to a wide variety of E. coli genes. The invention provides the sequences in such a way as to make them available for a variety of analyses. In one embodiment the nucleic acid sequences provided are present as an array of probes that may be used to measure gene expression of at least 20,000 E. coli genes and at least 500 intragenic regions. As such, the invention relates to diverse fields impacted by the nature of molecular interaction, including chemistry, biology, medicine, and medical diagnostics.

Chimeric proteins are expressed, secreted or released by a bacterium to immunize against or treat a parasite, infectious disease or malignancy. The delivery vector may also be attenuated, non-pathogenic, low pathogenic, or a probiotic bacterium. The chimeric proteins include chimeras of, e.g., phage coat and/or colicin proteins, bacterial toxins and/or enzymes, autotransporter peptides, lytic peptides, multimerization domains, and/or membrane transducing (ferry) peptides. The active portion of the immunogenic chimeric proteins can include antigens against a wide range of parasites and infectious agents, cancers, Alzheimer's and Huntington's diseases, and have enhanced activity when secreted or released by the bacteria, and/or have direct anti-parasite or infectious agent activity. The activity of the secreted proteins is further increased by co-expression of a protease inhibitor that prevents degradation of the effector peptides. Addition of an antibody binding or antibody-degrading protein further prevents the premature elimination of the vector and enhances the immune response.

The present invention relates to growing and testing microorganisms in a multitest format which utilizes a gel forming matrix for the rapid screening of clinical and environmental cultures. The present invention is suited for the characterization of commonly encountered microorganisms (e.g., E. coli, S. aureus, etc.), as well as commercially and industrially important organisms from various and diverse environments (e.g., the present invention is particularly suited for the growth and characterization of the actinomycetes and fungi). The present invention is also particularly suited for comparative analysis of phenotypic differences between cell types, including strains of microorganisms that have been designated as the same genus and species, as well as other cell types (e.g., mammalian, insect, and plant cells).

Provided herein is a novel E. coli O polysaccharide, O25B. Also provided herein are prokaryotic host cells comprising enzymes (e.g., glycosyltransferases) used in O25B production. The host cells provided herein produce O25B bioconjugates, wherein said bioconjugates comprise O25B linked to a carrier protein. Further provided herein are compositions, e.g., pharmaceutical compositions, comprising O25B and/or bioconjugates comprising O25B. Such compositions can be used as vaccines against infection with ExPEC, and may further comprise one or more additional bioconjugates.

A DNA construct that allows the inexpensive production of a target protein in E. coli, consisting of a nucleic acid sequence encoding a signal peptide which is operably linked with a gene coding for a carrier protein which is linked with a gene coding for the target protein via a gene coding for a cleavable sequence S, wherein the gene coding for a carrier protein is the spy gene from E. coli.

A reagent kit based on recombinant antigen for detecting Toxoplasma is prepared through taking 542-1218 fragment (t SAG1) from the primary surface antigen gene SAG1 of Toxoplasma, subcloning it to soluble expression carrier pET32a(+), transferring it to colibacillus, configuring engineering bacterium pET32a-tSAG1/BL21, IPTG induced efficient expression, ultrasonic splitting to obtain supernatant, purifying by Ni-NTA and Sephadex-G75, and coating the microholes on ELISA plate. Its test paper can also be prepared by same way.

The present invention discloses a performance improved recombination staphylococcus aureus protein A affinity ligand and a construction method thereof. The present invention adopts a molecular biology method. A sequence B of a nature protein A is selected for molecular transformation. A C-terminal of the sequence B is added with two cysteines, so that the protein A can pass through double-locus coupled chromatography matrix to stabilize the connection. Six glycines are added to the end of a second Loop of the sequence B to increase the length and reduce the binding force with an antibody, so that elution conditions are mild. On this basis, resistance performance to high concentration base of the protein A is transformed. Asparagines and phenylalanine at 23rd and 30th positions of the sequence B are respectively replaced by threonine and alanine to obtain a sequence Z with higher alkaline resistance properties. Then, isocaudarner is used for connecting sequence Zs of different numbers head-to-tail in series. The efficient expression system of e. coli is used for overexpression. The expressed recombination protein A is coupled to agarose matrix preparation affinity chromatography fillers and is used for purifying antibodies. Results show that the recombination protein A affinity ligand prepared by the present invention is good in elution performance and alkali resistance.

A bioconversion method of making vanillin including expressing VaoA gene in a mixture, expressing MtSAD1 gene in the mixture, feeding eugenol to the mixture, and converting ferulic acid to vanillin by incubating with a microbial Amycolalopsis sp. strain (Zhp06) and/or a recombinant E.coli strain.

The present invention relates to a preparation process of recombinant human parathyroid hormone PTH(1-34). The process constitutes a new engineering strain to result in high expression amount, simplepurifying course and low cost.The present invention adopts colibacillus degenerate codon modifying sequence corresponding cDNA, the basic sequence is L-V-P-R-PTH(1-34), where L-V-P-R is the enzyme incision site of thrombin. To the end of the nucleotides sequence, colibacillus terminator codon TAA is added; and in order to clone easily to the 5'-terminal enzyme incision site of EcoR I is added, and to the 3'-terminal, enzyme incision site of Sal I is added. High-purity recombinant huamn parathyroid hormone PTH(1-34) is obtained through fermentation of the engineering bacteria, occlusion body extraction, diluting renaturation, thrombase incision, anionic exchange chromatography and other steps.

The invention relates to a codon-optimized gene of a 16-type L2E7 recombinant protein of human papilloma virus (HPV), which can be expressed efficiently in colibacillus. The optimized gene can be expressed efficiently by being inserted in a prokaryotic expression vector pET9a, and the expression level accounts for 50% of all viruses; an expression protein is immune to mice after being purified. Proved by experiment results, the protein has the same immunogenicity with a protein which is not expressed by the optimized gene and can induce the release of specific gamma-1NF; proved by tumor growth inhibition animal experiments, protein immunity has evident inhibition effect on tumor growth and the growth of 90% of mouse tumor cells can be inhibited completely. The prokaryotic high-efficiency expression system of L2E7 recombinant protein, which is established by the optimized gene, can be used in the pilot scale production and drug discovery of preventive and curative vaccines of HPV16 infection and relevant cervical carcinoma.

An antimicrobial composition and a method for administering the antimicrobial composition through the water or feed of livestock, wherein the antimicrobial composition is made up of lysozyme and various other agents that act synergistically with lysozyme, such as, dried egg powder, albumen, a sequestering agent and/or a lantibiotic. The composition is used to inhibit the growth of, and diseases and epidemiological significant effects caused by, Clostridium perfinigens, E, coli and Salmonella, in the gut of livestock. More particularly, the antimicrobial composition and method relate to a feed additive that can be administered to poultry and/or swine through their feed. The use of such a feed additive may also inhibit other enteric pathogens that may be present in the gut of livestock.

The present invention discloses the preparation process of thynosin-alpha with acetylation modified N-end by using recombinant colibacillus. The preparation process includes obtaining thynosin-alpha gene, constructing recombinant expression vector containing thynosin-alpha, transforming prokaryotic cell, culturing prokaryotic cell and expressing thynosin-alpha, and separating and purifying thynosin-alpha with acetylation modified N-end. The present invention also discloses the process of utilizing recombinant colibacillus in preparing thynosin-alpha antigen with acetylation modified N-end and then cutting and separating to prepare thynosin-alpha-1 antigen with acetylation modified N-end, thynosin-alpha-2 antigen with acetylation modified N-end and similar matter. The thynosin-alpha with acetylation modified N-end has the functions of regulating immunity, cell proliferation, etc. and has wide application foreground in antivirus and antitumor.

The invention discloses a spirulina culture method, which comprises the following steps of (1) putting a solid culture medium into a sterilization pot to be steamed and boiled; (2) performing inoculation: spreading algae species of spirulina into the culture medium in a fully automatic pollution-free room; after the inoculation is completed, automatically and uniformly stirring the materials; (3) performing three-dimensional culture on the spirulina to obtain spirulina liquid; (4) performing harvesting, washing and concentrated dewatering; performing spray drying; completing the culture; (5) sieving the cultured materials to prepare the scaled culture spirulina. The spirulina culture method has the advantages that the germ content is low; the culture efficiency is high; high efficiency and health are realized; the energy consumption is low; the precipitation and the wall attachment of the spirulina are reduced; the collecting rate of the spirulina is improved; the scaled culture of the spirulina can be realized.

Genes for proteins which spontaneously form dimers and/or oligomers can be recombinantly linked together, which upon expression in E. coli produces stable dimeric fusion proteins that spontaneously self-assemble into enormous, polyvalent complexes having increased immunogenicity and functionality. Linear, network and agglomerate complexes with enormous sizes and polyvalences are constructed using glutathione S-transferase, Norovirus P domains (NoV P⁻ and NoV⁺), the protruding (P) domain of hepatitis E virus (HEV P), the astrovirus P domain (AstV), a monomeric peptide epitope (M2e of influenza virus), and/or a protein antigen (VP8* of rotavirus) fused in different combinations. The resulting complexes can contain hundreds to thousands NoV P-protein, HEV, AstV, M2e and/or VP8* copies and exhibit higher immunogenicity than the individual proteins alone. The large size and multivalent nature of the complexes are candidates as a bivalent or multivalent vaccines against Norovirus and other pathogens, and for generation of antibodies for diagnosis and research purposes.

The invention relates to a Bt protein in-vitro insect resistance identification method which comprises the following steps: connecting a Bt gene to a protocaryon expression vector pET-28B; transferring the protocaryon expression vector containing the Bt gene into colibacillus BL21; inducing to express the Bt protein; extracting the induction-expressed Bt protein; mixing the Bt protein into an Ostrinia nubilalis larva artificial feed, and feeding Ostrinia nubilalis larvae; and after several days, making statistics on the mortality and growth conditions of the Ostrinia nubilalis larvae. The method is simple and quick to operate, is not restricted by time and seasons, has the advantage of low economic cost, and can be used for quickly carrying out Bt protein insect resistance identification.

The invention relates to an HIV-1 integrase expression bacterial strain and an integrase inhibitor in-vitro screening model. In the model, HIV-1 integrase soluble expression plasmid and expression plasmid with integrase LTR sequence are firstly established based on gene engineering technology, and established recombinant plasmid pET28a-IN is transferred into commercial colibacillus BL21 to obtainthe integrase expression bacterial strain; and then the integrase expression bacterial strain is cultured in large-scale, finally integrase is separated and purified, works on the substrate plasmid and is added into samples to measure the activity. The detection model overcomes the defects such as high cost, high background and more influence factors in the existing ELISA detection method; the detection model can carry out primary screening on various natural or chemically synthetic integrase inhibitors in-vitro largely and quickly; and the detection model is an HIV-1 integrase in-vitro screening model with the advantages of quick operation, flexibility, fewer interference factors and low cost.

The present invention relates to the field of biotechnology. The invention provides a novel approach using tobacco mosaic virus omega leader sequence to enhance the solubility of the recombinant products in E. coli and the method of use therefore. The invention provides the utilization of tobacco mosaic virus omega leader sequence into E. coli expression vector, and the tobacco mosaic virus omega leader sequence containing expression vector can be used in combination with other available means to obtain higher expression or better solubility. The invention can be applied to biotechnological pharmaceutical industry, genetic engineering, biochemistry and molecular biology etc. The invention provides an expression vector pTORG, which is a highly efficient GST fusion expression vector, and can significantly enhance the yield of biologically active recombinant products.

The invention relates to a method for producing polypeptides or proteins in a two-chain form by recombinant expression in E. coli host cells, whereby i) the polypeptide or protein exerts its biological activity as a two-chain polypeptide or protein, ii) the C-terminal amino acid group of the first chain is an Arg or Lys group, iii) the second chain of the protein/polypeptide, at its N terminus, has 1 to 20 amino acid groups and a pentapeptide sequence VPXGS referred to as PRS, wherein X may be any naturally occurring amino acid, wherein V represents Val, Leu, Ile, Ala, Phe, Pro or Gly, P represents Pro, Leu, Ile, Ala, Phe, Val or Gly, G represents Gly, Leu, Ile, Ala, Pro, Phe or Val and S represents Ser, Tyr, Trp or Thr.