46results about How to "High expression activity" patented technology

The invention provides a method for culturing microzyme of secretory expression proteolytic enzyme. The method is characterized in that an inorganic salt culture medium is adopted for fermenting and culturing a recombinant methylotrophic yeast. Fermentation and culture comprise the growth period of the microzyme and the secretory expression period of the proteolytic enzyme. In the secretory expression period of the proteolytic enzyme, the feed flow of methanol can be controlled in real time according to the variation of oxygen dissolved in fermentation liquid, and the PH value of the fermentation liquid is adjusted with ammonia, thus expressing the proteolytic enzyme at high efficiency. Compared with the prior art, the method for culturing the microzyme of the secretory expression proteolytic enzyme has the characteristics of low culture cost, convenient and accurate reaction control mode and high expression activity of the proteolytic enzyme.

The invention discloses a construction method of double knockout recombinant rhodococcus. The construction method comprises the following steps: (1) primers are designed according to nitrile hydratase gene sequence of the rhodococcus for respective amplification of an upstream sequence and a downstream sequence of a nitrile hydratase gene, and an upstream fragment and a downstream fragment of the nitrile hydratase gene are obtained; (2) the upstream fragment and the downstream fragment of the nitrile hydratase gene are inserted into a suicide plasmid, and a recombinant suicide plasmid is obtained, wherein the suicide plasmid carries a kanamycin resistant gene and a levansucrase gene; (3) competent cells of recombinant rhodococcus of an amidase gene are knocked out through transformation of the recombinant suicide plasmid, and the double knockout recombinant rhodococcus is obtained: first screening is performed through kanamycin resistance, and a first recombinant colony is obtained and subjected to second screening through a sucrose plate. The invention further discloses recombinant rhodococcus with high nitrilase expression, a construction method of the recombinant rhodococcus and a method for preparing acrylamide.

The invention belongs to the field of plant genetic engineering and discloses an MYB type transcription factor GmMYB29 of Glycine max as well as an encoding gene and an application of the transcription factor GmMYB29. The transcription factor GmMYB29 has an amino acid sequence shown as SEQ ID NO.2. An open reading frame of the gene has a DNA sequence shown as SEQ ID NO.1. The transcription factor GmMYB29 increases the content of isoflavones of the Glycine max by promoting driving expression activity of a key structure gene promoter for an isoflavone biosynthetic pathway. The transcription factor GmMYB29 has significant value in breeding of a new Glycine max variety with high content of isoflavones.

The invention discloses a sea island cotton receptor albuminoid kinase gene promoter and an application thereof. The length of the promoter is 3183bp, nucleotide sequences are shown as SEQ ID NO.1. Receptor albuminoid kinase gene encoding protein products are relevant to the signal conduction of plant stress response, the gene promoter is the promoter with special vascular bundles and is expressed through biological or abiological inducement, the expression level of target genes in the vascular bundles by a gene engineering method through the promoter is enhanced, and the resistance of cotton on greensickness and abiotic stress is improved.

The invention discloses a technology for producing lycopene by biological fermentation and relates to the field of biological pharmacy. The technology for producing the lycopene by the biological fermentation, disclosed by the invention, comprises the following steps: inoculating positive fungus spores and negative fungus spores into saline solutions respectively to obtain a positive fungus sporesuspension solution and a negative fungus spore suspension solution; inoculating the positive fungus spore suspension solution and the negative fungus spore suspension solution into a seed culture medium according to the concentration ratio of 2 to (3 to 5) to obtain a seed solution; inoculating the seed solution into a fermentation culture medium and carrying out fermentation culture; after carrying out the fermentation culture for 12h, supplementing a fermentation culture medium which is equivalent to an original fermentation culture medium into a fermented solution. By adopting the technology disclosed by the invention, a form of the seed solution is improved; the positive spore suspension solution and the negative spore suspension solution are directly inoculated into the seed culturemedium to carry out conjugation; produced thalli are extremely small fungus pellets or thalli and pelletized thalli are not generated, so that the absorption of oxygen and nutrient substances in a liquid culture medium is facilitated; fermentation conditions are improved so that the metabolism of the thalli and the expression activity of a product are enhanced; the accumulation of the lycopene isfacilitated and the yield of the lycopene is improved.

The invention relates to a transgenic construct and application of the same in preparation of an epididymis head gene conditional knockout mouse model. Specifically speaking, it is discovered by the inventor that an Lcn5 gene promoter sequence with a length of 1.8 kb can drive expression of an exogenous gene in head (especially middle-and-far-end specific) areas of epididymis of a mouse and an exogenous Cre gene expression vector driven by the promoter is thus constructed. The transgenic vector can drive specific expression of an exogenous Cre gene at the head of the epididymis of an animal and expression of other genes like CreERT2 recombinase. Usage of such a transgenic system for preparation of a transgenic mouse, a high genomic integration rate (greater than 25%) can be obtained, and expression of an exogenous gene in medium-and-far-end areas of epididymis heads can be accurately and highly efficiently driven. The invention further establishes an epididymis head specific area conditional gene knockout or overexpression technology platform.

A method for determining whether a substance can increase the expression level of a fatty acid binding protein (FABP) in an animal. The method includes using a cell that includes an expression construct that comprises a FABP promoter operably linked to a polynucleotide sequence encoding a reporter molecule, wherein the cell is contacted with a candidate substance and then cultivating the cell under conditions conducive to expression of the reporter molecule. Increased expression of the construct in the presence of the candidate substance as compared to a control leads to improved sleep and long-term memory in the subject.

The invention discloses a method for reinforcing meat chicken cold stress resistance, and belongs to the technical field of agricultural cultivation. The method comprises the following steps: (1) illumination control, (2) chicken house environmental control, and (3) feeding control. According to the invention, the method can effectively guarantee normal development of meat chickens under cold stress conditions, improves the meat yield of the meat chickens and the development degree of intracorporeal organs, reduces the cultivation risk, and has extremely-good economic benefits and promotion value.

The invention discloses a method for improving the expression activity of aquaporin in a vesicle. The method comprises the following steps: 1, performing phosphorylation on the aquaporin and preparing a phospholipid vesicle system solution; and 2, introducing the phosphorylated aquaporin. The invention also provides an aquaporin reverse osmosis membrane. The aquaporin reverse osmosis membrane is prepared by using the phosphorylated aquaporin vesicle prepared by the above method. The expression activity of the aquaporin is improved through the phosphorylation of the aquaporin; and the phosphorylation modified aquaporin is embedded into the vesicle and applied to the reverse osmosis membrane, so under the condition of achieving water flux and desalinization rate with the same level, the ratio of the used aquaporin to the vesicle is greatly reduced.

The invention relates to the field of gene expression. The nucleotide sequence of a porcine alpha-interferon gene with efficient expression and high antiviral activity is shown as a sequence 2. Genes of the sequence 2 are cloned to a eukaryon expression vector to obtain a recombinant plasmid. A synthetic method of the porcine alpha-interferon gene comprises the following steps of: inserting a Rabbit beta-Globin Intron II, a pCMV immediate early promoter and a T7 promoter into pVAX1(C) to obtain pMVAX1(C); and inserting a maturation protein gene of pIFN-alpha into the pMVAX1(C) to obtain pMVAX1(C)-pIFN-alpha. The expression protein of the porcine alpha-interferon gene is applied to the preparation of a medicine for treating a porcine reproductive and respiratory syndrome. The porcine alpha-interferon gene with efficient expression and high antiviral activity breaks through the limitation that the expression amount is low and the activity is poor while being used for expressing the pIFN-alpha by the pMVAX1(C) carrier and the activity unit can be up to 2*10<7.0> U/0.1 mL; and through the porcine alpha-interferon gene provided by the invention, the proliferation of PRRSV (Porcine Reproductive and Respiratory Syndrome Viruses) with high pathogenicity can be obviously inhibited and the economic loss of a pig farm can be reduced.