197 results about "Cloned genes" patented technology

The present invention is a substantially purified sortase-transamidase enzyme from Gram-positive bacteria, such as Staphylococcus aureus. A specific sortase-transamidase enzyme disclosed has a molecular weight of about 29,076 daltons and catalyzes a reaction that covalently cross-links the carboxyl terminus of a protein having a sorting signal to the peptidoglycan of a Gram-positive bacterium, where the sorting signal has a a motif of NPQ / KTN / G therein. Variants of the enzyme, methods for cloning the gene encoding the enzyme and expressing the cloned gene, and methods of use of the enzyme, including for screening for antibiotics and for display of proteins or peptides on the surfaces of Gram-positive bacteria, are also disclosed.

The present invention provides an enzyme which catalyzes a reaction to transfer sugar to a hydroxyl group at position 2′ of chalcones and a gene thereof, and preferably an enzyme which catalyzes a reaction to transfer glucose to a hydroxyl group at position 2′ of the chalcones. Furthermore, the invention provides a plant whose flower color has been changed using the glycosyltransferase. Using probes corresponding to conservative regions of the glycosyltransferase, some tens of glycosyltransferase genes having nucleotide sequences corresponding to the conservative regions were cloned from flower petal cDNA libraries of carnation and the like. Furthermore, each of the glycosyltransferase genes was expressed in Escherichia coli, activity to transfer glucose to position 2′ of the chalcone, i.e., the glycosyltransferase activity at position 2′ of chalcone was confirmed in an extract solution of the Escherichia coli, and it was confirmed that cloned genes encoded the glycosyltransferase at position 2′.

A simple and highly efficient method for cloning cDNAs including CD27 (SEQ ID NO:28) from mammalian expression libraries based on transient expression in mammalian host cells has been discovered. Novel expression vectors allowing highly efficient construction of mammalian cDNA libraries are disclosed. The cloning method of the invention which has been used to clone genes for cell surface antigens of human lymphocytes, has general application in gene cloning. Cell surface antigens cloned according to the present invention have been purified, and the nucleotide and amino acid sequences determined. These antigens have diagnostic and therapeutic utility in immune-mediated infections in mammals, including humans.

The invention relates to methods of separation and / or purification of impurities yielding a purified heterologous protein product devoid of related impurities or with substantially minimal quantities of such glycosylated impurities. More specifically, the invention relates to the identification of glycosylated forms of insulin analogues such as glargine impurities characterized post expression in yeast based systems such as Pichia pastoris. The invention also relates to methods used to clone gene encoding the protein insulin glargine; inserting the related gene in a suitable yeast host; producing culture of the recombinant strain, stimulating expression of the heterologous polypeptide, its secretion and purification post fermentation and related enzymatic conversions.

The invention discloses a Yunnan red pear PyMYB gene through pigment synthesis regulation and trichome development regulatory protein as well as a prokaryotic expression vector thereof and an application of the prokaryotic expression vector. The PyMYB gene is cloned from red fruit peel of Yunnan red pear NO.1 by using a RT-PCR (reverse transcription-polymerase chain reaction) technology, and then the gene is expressed in escherichia coli Rosetta (DE3) by using the prokaryotic expression vector and is purified by a GST (Glutathione S-transferase) gel column purification method to obtain a Yunnan red pear PyMYB purified protein, wherein the Yunnan red pear PyMYB purified protein is used for the preparation of a PyMYB specific antibody and the application thereof in a PyMYB protein expression detection and immunoprecipitation, a chromatin co-immunoprecipitation and a fusion protein sedimentation experiment. The antibody prepared by the invention is strong in specificity, can accurately detect subcellular localization of the PyMYB protein, and is used for verifying protein interaction in the co-immunoprecipitation experiment, efficiently separating the protein and DNA (deoxyribonucleic acid) segment combined by the PyMYB protein and detecting the expression of the antibody in a transgenic plant.

The invention discloses a gene modification based method for expressing an exogenous drug through a probiotic and an application of the method, the homologous recombination and the one-step seamless cloning are used to knock out an indispensable gene dapA of escherichia coli Nissle1917 to obtain an escherichia coli nutritional deficiency strain Nissle1917deltadapA, the dapA gene is cloned to construct a complementary plasmid pMalc2x-dapA of the nutritional deficiency strain, the complementary plasmid pMalc2x-dapA is electrically shocked to convert into the strain Nissle1917deltadapA, an amicillin resistance gene ampR of a pMalc2x carrier is substituted through the homologous recombination to obtain a plasmid balance system without an antibiotics resistance marker, 12 amino polypeptide sequence genes of an amino terminal sall4 are synthesized in vitro at a sall4 amino terminal, a pelB signal peptide gene sequence of an erwinia carotovora pectinase is fused at the N end, a gene sequenceof a His protein tag is added at the C end, the gene sequences are closed into the complementary plasmid pMalc2x-dapA together to obtain an exogenous drug expression plasmid pMalc2x-dapA-sall4, the plasmid is imported into the escherichia coli nutritional deficiency strain Nissle1917deltadapA, the obtained strain effectively expresses the sall4 polypeptide, and the method has an obvious effect ontreating the liver cancer.

The invention relates to application of a herba saussureae involucratae sikCOR gene in cultivating a cold resistant plant. In the invention, a sikCOR gene is cloned from a herba saussureae involucratae, and a plant expression vector is constructed by utilizing the gene to obtain the cold resistant transgenic plant through genetic transformation. The sikCOR gene is cloned from the herba saussureae involucratae and then is expressed in the transgenic plant to improve the cold resistance of the transgenic plant, improve the low temperature stress resistance of the plant through excessive expression of the gene and finally obtain the plant with obviously enhanced low-temperature resistant capability. By screening and cloning genes directly related to cold resistance from the herba saussureae involucratae, the low-temperature adaptability mechanism of the saussureae involucratae is revealed. The herba saussureae involucratae sikCOR gene is used in breed improvement of crops, thus having great academic and economic value.

The invention provides an anti-citrus bacterial canker disease pthA-nls gene, a construction method and application thereof. The invention uses the pathogenic gene pthA-mediated broad-spectrum disease resistance theory of the citrus bacterial canker disease to construct a new anti-citrus bacterial canker disease gene (pthA-nls). The construction method comprises the following steps of: (1) primer design; (2) clone of NLS sequence; (3) recovery and enzyme digestion of PCR product; (4) double enzyme digestion of a plant expression vector; (5) connection between product of enzyme digestion of pBI121 and corresponding product of PCR enzyme digestion of the NLS sequence by T4 DNA ligase; (6) acquisition of positive cloned gene pthA-nls by converting ligation product into colon bacillus DH5alpha. The gene pthA-nls can further perform genetic transformation on susceptible varieties of orange so as to acquire new anti-citrus bacterial canker disease germ plasm. The anti-citrus bacterial canker disease pthA-nls gene provides an effective tool for breeding of anti-citrus bacterial canker disease.

The present invention relates to rice chlorine ion passage gene OsCLC, its coded protein and clone method. The gene has sequence indicated by SEQ NO 3. The gene of 2429bp is cloned from differentiated rice callus. The opening reading frame of cloned gene encodes protein of 808 amino acid (molecular weight 88KD)which has 11 trans-membrane region indicated by trans-membrane region analysis. Cluster analysis indicates the OsCLC gene is new member of CLC family. The OsCLC gene expression level has positive correlation with salt concentration rise in a certain time range and respond to the salt stress and other environmental stress. The gene can be induced to produce partial complementary GEF1 function in yeast mutant and partially inhibit the sensitivity to NaCl and recover growth function. The gene is sensitive to pH value variety and not sensitive to different cation.

The invention discloses an application of a corn transcription factor ZmMYB42 gene to drought-resistant breeding of plants. A gene ZmMYB42 is cloned from corn, the gene or a RNAi structure thereof isrecombined into a plant expression vector in a sense form, a fusion gene or a ZmMYB42RNAi structure is introduced into crops by using a transgenic technology, drought resistance determination is performed on transgenic plants, transgenic plants with obviously improved or reduced drought resistance and progenies thereof are screened out, and new plant germplasm with application value is created. The cDNA sequence of the corn transcription factor ZmMYB42 is as shown in SEQ ID No. 1, and the coded amino acid sequence of the corn transcription factor ZmMYB42 is as shown in SEQ ID No. 2. The plantsrefer to cultivated cereal crops. The application disclosed by the invention is of great significance for increasing the crop yield under drought stress.