34 results about "Chaperone Gene" patented technology

The invention relates to an unmarked gene-deleted and attenuated mutant strain of a vibrio alginolyticus wild strain, which is the attenuated active vaccine of the vibrio alginolyticus wild strain and in which the sRNA molecular chaperone gene hfq of the vibrio alginolyticus wild strain is deleted. The vibrio alginolyticus wild strain is EPGS020401, and the collection number of the vibrio alginolyticus wild strain is CCTCC NO.AB 209306. The invention also provides the preparation of the unmarked gene-deleted and attenuated mutant strain of the vibrio alginolyticus wild strain and the use thereof in prevention and control of vibrio alginolyticus disease in cultured fish. The unmarked gene-deleted and attenuated mutant strain of the vibrio alginolyticus wild strain or the preparation thereof, which are disclosed by the invention, eliminates universal potential environment and product safety risks of the conventional attenuated vaccine and is a safe, effective and economic vaccine for preventing and controlling vibrio alginolyticus diseases in cultured fish.

To provide an activator of gene expression of molecular chaperone gene and applications thereof.This activator of gene expression of molecular chaperone gene contains an eggshell membrane component, for example, a powder containing eggshell membrane or a soluble eggshell membrane component. Moreover, the powder containing eggshell membrane used in the present invention may be a fine powder preferably having 6 μm or less in the mean volume particle diameter of the fine powder containing eggshell membranes, and / or 20 μm or less in the maximum volume particle diameter thereof, but not limited to these values.

The invention discloses a method preparing a recombinant human nerve growth factor by using an Escherichia coli expression system. Beta-subunit genes of the recombinant human nerve growth factor comprise 6 histidine purification sites, an enterokinase cleavage site and a beta-subunit gene encoding mature peptide of modified human nerve growth factor and a molecular chaperone gene. The beta-subunit genes of the recombinant human nerve growth factor have high expression quantity in the Escherichia coli; generated inclusion body protein has high-efficiency renaturation under the assist of the molecular chaperone; and the recombinant human nerve growth factor obtained by nickel column affinity chromatography after the molecular chaperone is cleavaged accurately by the enterokinase, has a purity greater than 99% and biological activity greater than 500,000 AU / mg.

The invention discloses a primer, a probe, a kit and a detection method for detecting fusion mutations of human ALK gene, and the primer and probe include: designing on the broken exon sequences of six partner genes in the ALK fusion gene Multiple upstream primers, shared downstream primers and fluorescent detection probes designed on ALK exon 20; 6 partner gene break exon sequences in ALK fusion gene include: EML4‑exon2 / 6 / 13 / 14 / 17 / 20, TFG ‑exon5, KIF5B‑exon15 / 17 / 24, TPR‑exon15, HIPI‑exon21, SEC31A‑exon21; the kit can cover up to 28 ALK fusion forms, with high specificity and sensitivity, and can effectively avoid false positives. The established one-step reverse transcription detection method can simplify operation and avoid contamination.

The invention discloses an Escherichia coli expression system for producing oxalate oxidase, a production method of oxalate oxidase and application thereof. The recombinant expression plasmid in the Escherichia coli expression system comprises: an oxalate oxidase gene, a molecular chaperone gene and a gene for promoting manganese ion pumping into Escherichia coli cell-related proteins. The present invention obtains the optimal expression of oxalate oxidase by screening and testing the genes and copy numbers of different manganese ion-related proteins, the type of promoters controlling the expression of molecular chaperones, and the type of medium used in the production process of oxalate oxidase Combined, the expression activity of oxalate oxidase in Escherichia coli exceeds 80U / mL. The specific activity of the oxalate oxidase preliminarily purified from the supernatant of the cell disruption liquid is greater than 20 U / mg, and the molecular weight of the monomer is about 25 kDa. The technical scheme of the invention has the advantages of simple production and purification process, high expression amount and specific activity, easy industrial scale-up, low cost, and favorable industrial production and application of this type of enzyme.

The invention discloses an Escherichia coli expression system of recombinant protein containing manganese ions and an application method thereof. The recombinant expression plasmid in the Escherichia coli expression system is one of the following situations or a combination thereof: (1) at least comprising an Escherichia coli molecular chaperone gene and an enzyme protein gene containing manganese ions; (2) comprising an Escherichia coli molecular chaperone gene , enzyme protein genes containing manganese ions, and overexpression or inhibition of manganese ion channel-related protein genes; (3) including Escherichia coli molecular chaperone genes, enzyme protein genes containing manganese ions, and protein genes affecting intracellular manganese ion concentration . Through the construction and optimization of the Escherichia coli expression system, efficient soluble and active expression of the manganese ion-containing enzyme protein can be realized. Compared with the traditional method, the efficiency is high, the purification process is simple, and the cost is low, which is beneficial to the production of the manganese ion-containing enzyme. Industrial production and application.