35 results about "Phosphomannose Isomerase" patented technology

The disclosure provides new compounds and compositions thereof, and methods for treating or ameliorating a disorder relating to CDG-Ia. In particular, the disclosure provides benzoisothiazolone inhibitors of PMI, which have been synthesized and their ability to drive glycosylation has been demonstrated. The disclosure provides two synthetic routes for these compounds, including a new copper-catalyzed N-arylation reaction amenable to parallel derivitization. The disclosed compounds represent potent inhibitors of PMI, and their dose-dependent efficacy in cell-based models of glycosylation have been demonstrated. In addition, the disclosed compounds are selective over PMM and therefore, are useful in treating or ameliorating a disorder relating to CDG-Ia.

The invention belongs to the technical field of plant genetic engineering and relates to a method for screening a transgenic paddy plant by a phosphomannose-isomerase (PMI) gene of yeast. The method comprises the following steps of primer designing, PCR amplification, carrier construction, genetic transformation, screening and identification. A PMI gene obtained by cloning of brewer's eucaryon yeast is used as a selection marker. Through an agrobacterium-mediated transformation method, the PMI gene is introduced into a paddy acceptor. Transformed paddy callus is screened in mannose mediums having different concentration gradients so that a transgenic paddy plant is obtained. The method has the advantages that in plant transformation and screening, antibiotics and herbicides are not used; and the transgenic paddy plant screened by the method does not contain an antibiotic gene or a herbicide gene and is safe for the environment and a human body.

The invention discloses a gene and its protein of phosphomannose isomerase, and its application. The purpose is that providing a gene and its protein of phoshomannose isomerase, and applying the gene and protein in selecting trans-genic plant. The gene has one of nucleotide sequences as follow: 1) the SEQ ID NO: 1 DNA sequence in sequence table; 2) coding the DNA sequence of SEQ ID NO: 2 in the sequence table; 3) the nucleotide sequence that can hybrid with restrict DNA sequence of SEQ ID NO: 1 in the sequence table at high strict condition. Mannose has no harm for human and animal, degradation-liable in environment and selected marker system has the advantages of product yield safety, the selecting regent price is cheap, the selecting procedure is simplicity and the selected marker system don't impact the metabolism balance of transformation plant, the trans-genic plant grow vigorous, significance effect selection. The selected marker system has potential and extensive application outlook in trans-genic plant study and its market process; it may be the leading selected marker system in plant transformation.

The invention provides a method of effectively introducing exogenous genes into rice cells by utilizing PMI (phosphomannose isomerase) selective marker, a transformation system, as well as a preparation method of genetically modified rice by utilizing the method.

The invention provides a codon vegetalization-transformed PMI (phosphomannose isomerase) gene, which is designed and synthesized by utilizing optimized rice codon. Furthermore, the invention provides an expression cassette and an expression carrier, as well as applications of the expression cassette and an expression carrier on aspect of genetic transformation. A plant expression carrier is constructed by utilizing the transformed PMI gene, and an exogenous gene is injected to rice cells by taking mannose as a screening reagent. According to the codon vegetalization-transformed PMI gene, the high-efficiency genetic transformation of rice can be realized as transformation efficiency of the plant PMI subjected to codon vegetalization transformation is obviously improved.

The invention provides a phosphomannose isomerase gene OsPMI1 originated from oryza sativa. The nucleotide sequence of the phosphomannose isomerase gene OsPMI1 is as shown in SEQ ID NO:1. The invention further provides a prokaryotic expression vector containing the OsPMI1. The invention further provides an enzyme activity analysis method for the phosphomannose isomerase. Besides, the invention provides an expression box containing the OsPMI1 and a plant expression vector, as well as application of the expression box and the expression vector in the aspect of plant genetic transformation. According to the invention, the OsPMI1 gene is adopted to build the plant expression vector, mannose is used as a selective agent, and the transformation of oryza sativa cells is successfully realized; the phosphomannose isomerase gene originated from plant is successfully separated and cloned, no potential hazard is brought as the phosphomannose isomerase gene comes from plants (oryza sativa), and the phosphomannose isomerase gene originated from oryza sativa can be used for replacing the colibacillus-originated phosphomannose isomerase, so as to reduce the potential security risk.

The invention relates to a method for constructing recombinant escherichia coli, synthesizing GDP-fucose by using mannose, further producing 2 '-fucosyllactose and improving the utilization rate of mannose, and belongs to the field of microbial metabolism engineering. The engineering bacterium takes escherichia coli as a starting strain, a Plac promoter sequence and lacI and lacZ genes in a lac operon sequence are knocked out, wcaG, gmd and lacy genes are overexpressed on 1acI and lacZ gene loci, then a phosphomannose isomerase coding gene manA is knocked out on a genome, a phosphomannose mutase coding gene manB and alpha-(1, 2, 3, 4, 5, 6, 7, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8 and (2) obtaining a fucosyl transferase coding gene futC and a mannose-1-phosphate guanine transferase coding gene manC. According to the fermentation strategy for producing the 2 '-fucosyllactose through the de novo synthesis route of the 2'-fucosyllactose, the utilization rate of a carbon source for producing the 2 '-fucosyllactose through escherichia coli fermentation can be greatly increased.

The invention provides a method for introducing a foreign gene into cleistogamy japonica rice by utilizing a PMI (phosphomannose isomerase) selection marker. Specifically, the method is characterized in that the foreign gene is introduced into the cleistogamy japonica rice variety 8m30 through mediation of an agrobacterium bacterial strain EHA 105 containing a pCAMBIA1381-PMI vector in the presence of mannose serving as a selection agent. PCR (polymerase chain reaction) detection and identification show that the foreign gene is introduced into the cleistogamy japonica rice variety 8m30. The method for introducing the foreign gene into the cleistogamy japonica rice by utilizing the PMI selection marker has the advantages that genetic transformation of the cleistogamy japonica rice variety is successfully realized, no antibiotics or herbicide is needed for selection, a character improvement process is sped up, gene drift caused by outward propagation of pollen is avoided, and further environment friendliness is improved.

The embodiment of the invention discloses a biological enzyme synthesis method and application of a cosmetic-grade hexose-6-phosphoric acid composition, and the method comprises the following steps: transforming a coding gene 17 of polyphosphate dependent kinase into escherichia coli to construct a genetically engineered bacterium I; the method comprises the following steps: transforming a coding gene BSMPI of phosphomannose isomerase into bacillus subtilis, and constructing a genetically engineered bacterium II; carrying out combined fermentation on the genetically engineered bacterium I and the genetically engineered bacterium II to prepare a crude enzyme solution of the compound enzyme; and carrying out enzyme catalytic reaction on the crude enzyme liquid of the compound enzyme to prepare the cosmetic-grade hexose-6-phosphoric acid composition. According to the method, biological enzyme does not need to be purified, and an enzyme catalysis product does not need to be refined and dried, so that the production and application cost is greatly reduced; in addition, cell experiments, skin tests and other means prove that the hexose-6-phosphoric acid composition disclosed by the invention has good effects of improving cell energy metabolism efficiency, promoting skin repair and regeneration, relieving skin wrinkles and the like.

The invention provides a phosphomannose isomerase gene ChlaPMI with the plant origin. The nucleotide sequence of the phosphomannose isomerase gene ChlaPMI is shown as SEQ ID NO: 1. The invention further provides a prokaryotic expression vector with ChlaPMI, and the prokaryotic expression vector can be used for identifying the ChlaPMI proteometabolism mannose activity. Based on the prokaryotic expression vector, the ChlaPMI proteometabolism mannose activity is identified so that it can be identified that the prokaryotic expression vector can carry out metabolism on the mannose. The invention provides an expression box with the ChlaPMI, a plant expression vector and application of expression box and expression vector in the plant genetic transformation application. The plant expression vector established through the ChlaPMI gene is utilized, the mannose serves as the screening reagent, and rice cells are successfully converted. The phosphomannose isomerase gene with the plant origin is successfully separated and cloned, and due to the fact that the phosphomannose isomerase gene comes from the plant (Chlamydomonas reinhardtii), the phosphomannose isomerase gene is an environmentally-friendly natural substance, and potential danger cannot be caused to the human beings. The phosphomannose isomerase gene can be used for replacing phosphomannose isomerase from the Escherichia coli.

The object is to develop a bacterium capable of fermenting glucose, mannose and xylose simultaneously, which can ferment a saccharified solution of a cellulose-type or lignocellulose-type biomass resource to produce ethanol, and to construct an energy-saving high-efficiency bioethanol conversion process. Thus, disclosed is Zymomonas mobilis bacterium which is prepared by integrating a gene encoding a phosphomannose isomerase derived from Escharichia coli into a levansucrase gene located on the chromosome by the double cross-over by means of a homologous recombsination method, and then introducing recombinant DNA prepared by binding a DNA fragment containing genes encoding a xylose isomerase, a xylulokinase, a transaldolase and a transketolase, respectively, all derived from Escherichia coli to a vector. Also disclosed is a method for producing ethanol by continuously fermenting a saccharified solution of a cellulose-type biomass resource in a system on which the Zymomonas mobilis bacterium is immobilized.

The invention discloses a monoclonal antibody used for specifically recognizing a non-antibiotic safe screening marker PMI in a transgenic plant and a preparation method thereof. The objective of the invention is to provide a key reagent material for immunological detection of the screening marker PMI in a natural transgenic species (corn, paddy rice, cotton, etc.) A PMI gene is a biologically safe marker gene. An antigen for preparation of the antibody is a soluble full-length PMI recombinant protein expressed by Escherichia coli; the eventually obtained antibody belongs to IgG1 subtype; and a sequence coding the variable region of the antibody is obtained through gene cloning. According to results of western blotting detection of a natural transgenic paddy rice material, the antibody has good specificity and can be used for detection of a transgenic material.

The present invention belongs to the field of plant transgenic technology, and discloses a method for improving sugarcane genetic transformation efficiency by taking mannose as a screening agent. The method is based on sugarcane young heart leaves-induced yellowish and granular type II embryogenic callus as a transgenic acceptor material, employs a negative pressure assistant and improved agrobacterium tumefaciens mediated method to lead a 6-phosphate mannose isomerase gene into a sugarcane genome, and enables getting transgenic sugarcane plants after screening and differentiating on mannose-containing medium. The method uses a PMI positive selection marker system, requires a short cycle from sugarcane callusing to differentiating into seedlings, is stable in transformation effects, and helps to overcome the defects that a genetic transformation system taking antibiotics and herbicides as negative selection markers is low in transformation efficiency, long time in embryogenic callus induced differentiation, easy in sugarcane callus browning, etc. The transformed callus maintains a high regenerative capacity, and has a transformation rate of 58.97% showed by molecular detection results.

The invention provides a phosphate mannose isomerase (PMI) gene OsPMI3 originating from a plant. A nucleotide sequence of the PMI gene OsPMI3 is obtained after carrying out addition, deletion, replacement or insertion of one or a plurality of nucleotides on a nucleotide sequence shown in SEQ ID NO:1. The invention also provides a prokaryotic expression vector containing OsPMI3 and an enzyme activity analysis method of PMI. Oryza sativa cell transformation is successfully achieved by utilizing the constructed plant expression vector of the gene OsPMI3 and taking mannose as a screening agent. The PMI gene originating from the plant is successfully separated and cloned. Originating from the plant (oryza sativa), the PMI gene does not pose a potential hazard to human. The PMI gene can be used to replace PMI from escherichia coli, thus reducing potential safety risks.

The invention provides a phosphate mannose isomerase (PMI) gene OsPMI2 originating from a plant. The PMI gene OsPMI2 is separated and cloned from oryza sativa. A nucleotide sequence of the PMI gene OsPMI2 is shown in SEQ ID NO:1. The invention also provides a prokaryotic expression vector containing OsPMI2, an enzyme activity analysis method of PMI, an expression cassette containing OsPMI2, a plant expression vector and an application of the expression cassette and the expression vector to plant genetic transformation. Oryza sativa cell transformation is successfully achieved by utilizing the constructed plant expression vector of the gene OsPMI2 and taking mannose as a screening agent. The PMI gene originating from the plant is successfully separated and cloned. Originating from the plant (oryza sativa), the PMI gene does not pose a potential hazard to human. The PMI gene can be used to replace PMI from escherichia coli, thus reducing potential safety risks.

The invention relates to a preparing method of a double-antibody sandwich enzyme linked immunosorbent assay (ELISA) kit detecting the content of phosphomannose isomerase (PMI) in transgenic plant and uses of the kit. The objective of the invention is to provide a detecting method for detecting whether a non-antibiotic selection marker that is the PMI exits in transgenic plant (corn, rice, cotton, and the like) or not and for quantifying the content of the PMI in the plant. A monoclonal antibody used in the detecting method is specifically described in the patent for invention 201410387508.1. ELISA detection on transgenic rice materials shows that the detecting method is high in sensitivity, high in specificity, simple and convenient in operation, rapid in qualitative detection and accurate in quantification, the lowest detectable limit for the PMI is 1.18 ng/mL, the linear detection range is 21-560 ng/mL, and the detecting method can be used for detection of transgenic materials.