73 results about "Small subunit" patented technology

A promoter isolated from a cotton gene encoding the small subunit of ribulose bisphosphate carboxylase is described. The isolated promoter is operably linked to a coding sequence of interest to make a chimeric gene.

The invention discloses a gateway cloning technology entry plasmid vector. The vector contains a photoinducible promoter (Prbcs) of a small subunit gene of 1,5-bisphosphate carboxylase (Rubisco) and a green fluorescent protein (GFP) reporter gene. The invention also discloses a construction method and application of the vector. Through the vector, the gateway cloning technology can be utilized to quickly construct a photoinducible plant expression vector of a target gene to realize the high level expression of the target gene in plant leaves, the expression of the target gene is adjusted by light intensity, and expressed target protein can be positioned into chloroplast or cytoplasm.

The invention belongs to the technical field of bioanalysis, and particularly relates to a method for analyzing microbial community structures. The method comprises the following steps of: extracting total RNA in an environment sample; separating small-subunit rRNA from the total RNA; carrying out reverse transcription on the small-subunit rRNA by random primer for sequencing; extracting the small-subunit rRNA sequence from the results of sequencing; and confirming category message according to the obtained small-subunit rRNA sequence, and acquiring the characteristics of the microbial community structure. The method does not use specific PCR amplification so as to avoid various deviations in the PCR process; and meanwhile, the method can be used for analyzing the community structure of bacteria, archimycetes and eukaryotic microorganism, thus truly and completely reflecting the characteristics of the microbial community structure in the environment sample.

The invention discloses a rice protein capable of promoting chloroplast development and a coding gene and application thereof. The protein is one of 1) a protein formed by amino acid residue sequences of the SEQ ID No.2 in a sequence table or 2) a protein which is formed after one or more amino acid residues in the amino acid residue sequences of the SEQ ID No.2 in the sequence table are substituted and/or deleted and/or added and has the same activities as the amino acid residue sequences of the SEQ ID No.2. The gene is used for coding the small subunit of ribonucleotide reductase, and the development of rice chloroplast is affected after the gene is mutated so that white stripes are distributed on the rice leaves in the vein direction.

The invention in particular relates to a plant expression vector pH2-35S-PrbcS-AMDH for improving the aluminum toxicity resistance of plants, a construction method and application thereof, which belong to the field of plant gene engineering. The special vector pH2-35S-PrbcS-AMDH for improving the aluminum toxicity resistance of the plants is the plant expression vector containing a photoinducible promoter (PrbcS) of a rubulose-1, 5-bisphosphate carboxylase (RubIsco) small subunit gene and an arabidopsis thaliana cytosolic malate dehydrogenase gene (AMDH). The AMDH gene is cloned from arabidopsis thaliana, the photoinducible promoter is used to control the overexpression of the AMDH gene in tobacco, malic acid is synthesized, and the malic acid is secreted out of cells so as to strengthen the resistance of the plants on aluminum toxicity in acid soil. Experimental results show that the activity of malate dehydrogenase of trans-AMDH genic tobacco leaves is 1.4 times of that of wild tobacco. Under the stress of 30 mu M of aluminum toxicity, trans-AMDH genic tobacco can secrete more organic acid, and has better root system growth; and the growth condition under the stress of the aluminum toxicity shows that the plant height and the green leaf number of the trans-AMDH genic tobacco are higher than those of the wild tobacco.

The invention discloses a xanthine dehydrogenase intercepting body and an application thereof. The protein comprises xanthine dehydrogenase intercepting body small subunit, xanthine dehydrogenase intercepting body intermediate subunit, and xanthine dehydrogenase intercepting body large subunit. The affinity of xanthine dehydrogenase intercepting body is increased by 19% by comparing a wild substrate (xanthine), the conversion number is increased by 115%, the catalysis efficiency is increased by 166%, and the temperature toleration is increased by 11 DEG C. The xanthine dehydrogenase intercepting body is in favor of developing the novel application field by combining with other enzymes, and is suitable for sample detection for detecting xanthine and hypoxanthine, the xanthine dehydrogenase intercepting body can be used for detecting inorganic phosphoric acid by combining with PNP enzyme, can be used for detecting adenosine deaminase by combining with PNP, XOD and POD enzymes and detecting 5'-nucleotidase, and is suitable for industrial application. The xanthine dehydrogenase intercepting body is in favor of increasing enzyme catalysis efficiency and reducing cost, and is in favor of industrial application.

The subject invention concerns materials and methods for providing plants or plant tissue with increased resistance to heat conditions. Increased resistance of a plant or plant tissue to heat conditions provides for decreased yield losses generally observed at elevated temperatures. One aspect of the invention concerns polynucleotides that encode a mutant plant small subunit of AGPase. The subject invention also comprises a mutant plant small subunit of AGPase encoded by a polynucleotide of the invention. The subject invention also concerns plants comprising a polynucleotide of the invention and method for making the plants.

The invention discloses a recombined streptomycete, a construction method of the recombined streptomycete and a method for increasing the antibiotic yield. The recombined streptomycete over-expresses acetolactic acid synthase small subunit ilvH. The recombined streptomycete over-expresses acetolactic acid synthase to adjust subunit genes, and the yield of products such as antibiotics for conducting secondary metabolism with L-isoleucine and/or L-valine and/or L-leucine as precursors/a precursor can be remarkably increased.

The subject invention concerns polynucleotides encoding a small subunit of plant AGP having one or more mutations in the amino acid sequence wherein the mutation confers increased heat stability to the expressed AGP enzyme. Mutations in the N-terminus of the small subunit of heat labile plant AGP results in AGP enzymes that are significantly more heat stable compared to wild type AGP in that the mutant AGP retains significant levels of enzymatic activity following exposure to heat treatment. In one embodiment, the polynucleotide encodes a mutant small subunit of maize AGP. The subject invention also concerns methods for providing a plant with increased resistance to heat conditions. Plants with heat labile AGP can be transformed with a polynucleotide of the present invention. The subject invention also concerns these transformed plants and transgenic progeny thereof. The subject invention also concerns mutant polypeptides encoded by polynucleotides of the present invention.

The invention relates to related protein capable of adjusting translation efficiency of chloroplast protein and improving heat resistance of plants, and application thereof. In particular, the inventor screens one mutant rps1 related to the translation efficiency of the chloroplast protein and the heat resistance of the plants from various different arabidopsis mutants, and identifies a plant chloroplast ribosome small subunit S1 (RPS1) gene from the mutant; and the gene and polypeptide can effectively maintain the heat stability of a thylakoid membrane, can improve the translation efficiency of the chloroplast protein of the plants, can improve the survival ability and the photosynthesis efficiency of the plants under heat stress, and can effectively avoid and reduce adverse influence of high temperature on the plants.

The invention relates to a gateway inlet vector for the gateway technology, and in particular to a gateway inlet plasmid vector (pEn-L4*-PrbcS-*T-GFP-L3*). The vector comprises sequences L4 and L3, rubisco, a small subunit gene photo-inductive promoter (PrbcS), a chloroplast stroma positioning sequence (T*) and a green fluorescent protein (GFP) reporter gene, which are required for the LR recombination reaction in the gateway technology. A construction method and application of the vector are that: one plant expression vector connected in series with two target gene expression boxes can be quickly constructed by using the vector and another gateway inlet vector containing L1 and L2 sequences through the LR recombination reaction so as to realize conversion operation of two target genes through one conversion event. The expression of the target gene can be controlled with the PrbcS promoter in the vector and high-level expression of the target gene in the leaves of a plant can be realized by substituting the GFP gene in the vector with the target gene.

The invention provides an ADP-glucose pyrophosphorylase mutant and a screening method and application thereof. The screening method includes the steps of firstly, synthesizing first chain cDNA; secondly, performing PCR amplification; thirdly, building AGPase large and small subunit cDNA prokaryotic expression vectors; fourthly, performing active screening. The method has the advantages that distant hybridization is performed on AGPase large subunit genes and small subunit genes from different species, and the recombinant gene expression vectors are obtained in a highly-controllable manner; escherichia coli glgC mutant strains are converted and inoculated to a Conberg enrichment solid culture medium, and iodine-potassium iodide dyeing is used to screen the high-activity ADP-glucose pyrophosphorylase mutant. The screening method is simple and practical and stable and reliable. The enzyme activity and glycogen content of the ADP-glucose pyrophosphorylase mutant are evidently higher than those of wild corn AGPase, evident change of the substrate affinity of the ADP-glucose pyrophosphorylase mutant is avoided, and the sensitivity of the ADP-glucose pyrophosphorylase mutant to activating agent is increased.

The invention discloses a maize-endosperm ADP-glucose pyrophosphorylas mutant and sieving method and application, which comprises the following steps: 1) mutating cDNA of large and small subunit of maize-endosperm AGPase through alcaine hydroxylamine mutation agent randomly; 2) transmitting mutated cDNA and wild-type cDNA into escherichia coli glgC mutation germ; sieving positive monoclonal; 3) seeding positive monoclonal line on the Conberg rich solid board to culture under 35-39 deg.c; 4) dyeing; comparing the color; obtaining deep color mutation as maize-endosperm ADP-glucose pyrophosphorylas mutant.

The invention belongs to the technical field of biology, and discloses an electrophoresis method for removing Rubisco from plant leaves and enriching and separating residual proteins. The method comprises the following steps of: removing Rubisco from watermelon leaves by using 18 percent PEG-4000; leaching residual proteins through a TCA-acetone precipitation method; and performing two-dimensional electrophoresis to separate residual proteins from watermelon leaves. Compared with a holoprotein two-dimensional electrophoretogram, the method has the advantages that: after Rubisco in watermelon leaves is precipitated by adopting the 18 percent PEG, most large and small subunits of Rubisco disappear, the expression levels of most residual proteins are raised, the quantity of proteins obtained by separating is increased by 48.4 percent, and a large quantity of new protein spots (65+/-15) appear in a region of which the molecular weight is smaller than 25.0kDa. As proved by repeated experiments, the electrophoresis method has the advantages of high repeatability, clear spectra and reliable result.

The invention discloses a unit splitting modulation method for focusing light through a scattering medium and relates to the field of bioimaging. The method is characterized by including the step of finding an optimal phase and the step of quaternary splitting method modulation. In the step of finding the optimal phase, the phase change of 0-2pi is conducted on a spatial light modulator, phase modulation is conducted on incident plane waves, a CCD is adopted to receive a corresponding speckle pattern, the light intensity at the focusing position is calculated, comparison is conducted, and the phase value corresponding to the maximum light intensity is stored; In the step of quaternary splitting method modulation, a phase (optimal phase) making the output strength at the focusing position maximum is found in each unit, then the units are equally and sequentially divided into four parts, each unit adopts a previous optimized phase while continuing to search for better phase distribution, and in this way, the units can be subdivided and optimized into smaller subunits, even pixels of the spatial light modulator.

The invention discloses a plant expression vector pK2-35S-PrbcS-*T-hps/phi for improving the metabolic capability of assimilating and formaldehyde absorption of plants, and the plant expression vector is a plant expression vector of a bifunctional enzyme HPS/PHI gene formed by integrating a PrbcS containing a Rubisco 3C small subunit gene and a transfer peptide sequence thereof as well as 6-phosphonate hexulose synthetase and 6-osphofructose isomerase of Mycobacteriumgastri MB19. A procaryon expression vector pET-23a-hps/phi plasmid of HPS/PHI is used as a template and is amplified to acquire an hps/phi gene, and the PrbcS and a chloroplast transfer peptide are used for controlling the expression of the hps/phi gene in the chloroplast of plant leaves, thereby improving the capability of assimilating and formaldehyde absorption of the plants. An experiment indicates that the growing condition of transgenic plants is better than that of wild plants, and the transgenic plants have a lighter phenomenon of leaf chlorosis when the transgenic plants grow in the environments of high-concentration extrinsic gas and liquid formaldehyde. Transgenic plant leaves can completely eliminate the formaldehyde in 4mM of formaldehyde treatment fluid within 90 hours, but wild leaves have slower absorption efficiency, and 17 percent of formaldehyde still remains.

The invention belongs to the technical field of biology and in particular relates to a recombinant transient expression vector carrying a small subunit gene of tobacco ribulose-1,5-bisphosphate carboxylase. One transient expression vector NtRBSC1-pFF19-GFP (Green Fluorescent Protein) is constructed by utilizing a tobacco NtRBSC1 gene; after tobacco cells are transformed through the vector, fused protein of fluorescence labeled gene GFP and NtRBSC1 can be transformed into tobacco protoplasts, so that NtRBSC1 protein is subjected to sub-cellular localization by detecting the position of the fused protein through a laser confocal microscope; the position of chloroplast is displayed. Overall, the manner is a simple and accurate method for localizing in living cells and accurate reference evidence can be provided for research on the chloroplast of the tobacco cells and related genes; research on the tobacco NtRBSC1 gene and deep development of functions of the tobacco NtRBSC1 gene are convenient to realize.

This invention provides an alternative source of ahas and pds nucleic acids for plant transformation and selection. In particular, the invention provides ahas and pds nucleic acids from cyanobacteria, for example, Synechocystis, and expression elements of these genes for control of expression in plastids. The invention further provides nucleic acids encoding the acetolactate synthase (ahas) large subunit and the ahas small subunit which were found to provide herbicide resistance to plants. The present invention also provides a novel Synechocystis mutant phytoene desaturase (PDS) gene conferring resistance to 4′-fluoro-6-[(alpha, alpha, alpha,-trfluoro-m-tolyl)oxy]-picolinamide, a bleaching herbicide. The present invention provides improved methods involving cyanobacteria for the screening of compounds, including a new high throughput protocol that is a rapid and cost effective way to identify target site genes.

The present invention provides a method of mapping and identifying new sites for antibiotic action in the ribosome of a microorganism. The method comprises: (a) providing a random mutant library of the rRNA genes of the microorganism prepared by randomly mutating the rRNA genes of the microorganism; (b) enriching the library in clones with deleterious rRNA mutants by negative selection; (c) screening for clones with deleterious rRNA mutations; (d) mapping the deleterious rRNA mutations in the clones obtained from step (c) to identify sites in the rRNA which are important functional sites in the ribosome; and (e) selecting functional sites identified in the rRNA in step (d) which are not targeted by a known antibiotic as new sites for antibiotic action for the microorganism. The rRNA gene can be the 16S rRNA of the small subunit or the 23S rRNA or the 5S rRNA of the large subunit of the ribosome of the microorganism.

Various genes of herpes virus of turkeys (HVT), Marek's disease virus (MDV) and infectious laryngotracheitis virus (ILTV) have been identified as non-essential regions (and candidates for insertion sites for foreign genes) and/or as antigen-encoding regions. The former include the HVT homologue of the HSV (herpes simplex virus) gC gene, the TX (thymidine kinase) region of MDV or ILTV, ORF3 of ILTV (as defined herein), the ribonucleotide reductase (large subunit) gene of ILTV, MDV or HVT and the ribonucleotide reductase (small subunit) gene of MDV. The antigen-encoding regions include the HVT homologues of the HSV gB, gC and gH genes, the ILTV homologue of HSV gB, ORF2 of ILTV, and the HVT homologue of the HSV-1 immediate early genes IE-175 and IE-68. Manipulation of these genes allows vaccines to be prepared comprising attenuated virus or virus carrying heterologous antigen-encoding sequences.

The invention relates to engineering bacteria based on nitrite reductase and an implementation method of the engineering bacteria in the technical field of genetic engineering. The method comprises the following steps: by taking streptomyces griseorubens genome DNA as a template, performing PCR amplification with a primer containing enzyme cutting sites to sequentially obtain nucleotide sequences for encoding nitrite reductase large-subunits and nitrite reductase small subunits; sequentially connecting the gene sequences obtained by amplification to a coexpression vector pETDuet-1, and finally obtaining a recombinant coexpression vector pETDuet-NC-ND; and transforming the nitrite reductase expression vector into an Escherichia coli expression strain. Aiming at the defects that the application range and effect are severely limited because most of the nitrite reductases are intracellular enzymes in a living body and the expression quantity is low in the prior art, the engineering bacteria can realize great in-vitro expression synthesis of the nitrite reductase by using genetic engineering means and has high significance for rapidly repairing secondary salinization soil and realizing precision agriculture.