3522 results about "Wild type" patented technology

The present disclosure provides guided microbial remodeling (GMR) methods for the rational improvement of plant-associated microbes to perform plant-beneficial functions. The GMR methods described herein allow for non-intergeneric genetic optimization of key regulatory networks within the microbes, which improve plant-beneficial functions over wild-type microbes but don't have the risks associated with transgenic approaches (e.g., unpredictable gene function, public and regulatory concerns, etc.). The present disclosure also provides remodeled microbes and compositions thereof. The utilization of remodeled microbes and compositions thereof will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived fertilizers.

The invention belongs to the technical field of gene engineering, and particularly relates to applications of OpgG and a mutant thereof in production of human milk oligosaccharide. The invention provides a dextran biosynthetic protein OpgG mutant, and the OpgG mutant is obtained by mutating the 443rd amino acid from threonine to proline on the basis of a wild type dextran biosynthetic protein OpgG as shown in SEQ ID NO.1. The invention also provides a preparation method of the dextran biosynthetic protein OpgG mutant. An opgG gene and / or a T443P mutant coding gene are / is introduced into a human milk oligosaccharide production strain and applied to production of human milk oligosaccharide, and the yield of 3 '-SL, 6'-SL, LNT II, LNnT and LNT is greatly increased.

The invention belongs to the technical field of biology, and particularly provides UCRT X heat-resistant reverse transcriptase mutants with improved thermal stability, the UCRT heat-resistant reverse transcriptase mutants provided by the invention comprise four single-point mutants, six double mutants and four three mutants, and compared with wild-type UCRT X heat-resistant reverse transcriptase, the mutant has longer half-life period at 65 DEG C; the double mutant has a better effect, and the half-life period of the double mutant is about four times of that of wild type DNA polymerase. The invention also provides a genetically engineered bacterium containing the mutant. The UCRT heat-resistant reverse transcriptase mutant obtained by the construction method provided by the invention has better thermal stability, shows higher thermal stability when being used for synthesizing DNA (Deoxyribonucleic Acid) by reverse transcription at higher temperature, and has higher application potential.

According to the barley HvPHR1 gene and the application thereof in the aspect of increasing the phosphorus content of barley, rice PHR2 homologous protein serves as bait, homologous comparison means is utilized, it is found that two PHR proteins HvPHR1 and HvPHR2 exist in barley, and it is found that the content of inorganic phosphorus in barley leaves can be remarkably increased through overexpression of the HvPHR1 gene in an HvPHR1 overexpression strain with the normal phosphorus concentration being 200 micrometers NH4H2PO4 and an HvPHR1 overexpression strain with the low phosphorus concentration being 10 micrometers NH4H2PO4 hydroponic culture for three weeks and a wild type control (GP). In conclusion, the function of the HvPHR1 gene in barley low phosphorus stress is defined, and a theoretical basis is provided for cultivation of new phosphorus-efficient barley varieties.

The invention discloses a high temperature resistant xanthan gum lyase mutant and a preparation method thereof, wild type xanthan gum lyase comes from Paenibacillus anensis, site-directed mutagenesis is carried out on the wild type xanthan gum lyase to obtain the high temperature resistant xanthan gum lyase mutant, and the mutant comprises at least one of N66V, R292A and S676V. The enzymatic activity of the xanthan gum lyase mutant treated in a buffer solution with the pH value of 5.0 at the temperature of 65-85 DEG C for 2 h is remarkably improved compared with that of an original enzyme, and the enzymatic activity of N66V / R292A / S676V is improved by 3.8 times compared with that of the original enzyme at the temperature of 85 DEG C.

Provided herein are gene editing pairs comprising an engineered protein, e.g, an engineered CRISPR protein and a guide ribonucleic acid (gRNA) that may provide improved gene editing relative to reference, i.e., wild type sequences. In some embodiments, the engineered protein of the gene editing pair comprises a RuvC cleavage domain with one or more amino acid modifications relative to a reference CRISPR protein RuvC sequence. In some embodiments, a guide ribonucleic acid (gRNA) of the gene editing pair comprises at least one modification in a scaffold stem loop relative to a reference scaffold stem loop sequence.

The invention discloses a hemoglobin-resistant Taq DNA polymerase mutant and a construction method thereof, and relates to the field of biology, and the hemoglobin-resistant Taq DNA polymerase mutant is characterized in that a nucleotide sequence for coding the Taq DNA polymerase is shown as SEQ ID NO.1, and the mutant comprises at least one mutation or all mutations selected from S623D and E721A sites. On the basis of natural Taq DNA polymerase, the molecular structure of the Taq DNA polymerase is modified through rational design and combination with a site-specific mutagenesis biotechnology, so that the Taq DNA polymerase mutant is more suitable for PCR amplification of samples containing hemoglobin and the like than a wild type, and the result judgment accuracy of a sample containing a hemoglobin inhibitor is more facilitated.

The invention discloses application of an OsNLP3 gene, and relates to the technical field of plant genetic engineering, and an amino acid sequence coded by the OsNLP3 gene is shown as SEQ ID NO.2. By constructing an OsNLP3 gene overexpression plant and comparing the OsNLP3 gene overexpression plant with a wild type of the OsNLP3 gene overexpression plant, the increase of the single gene expression quantity of the OsNLP3 gene overexpression plant causes the significant increase of the total protein content of the rice, the invariable characters of the grain length, the grain width, the yield and the like, the genetic effect is significant, and the OsNLP3 gene overexpression plant has huge application potential and prospects for the improvement of the characters of rice varieties and provides a new gene resource for

The invention includes a mutant T7 DNA ligase or a biologically active fragment thereof, which has greater activity than wild type T7 DNA ligase. The mutant T7 DNA ligase, or the biologically active fragment, has one or more substitutions differing from the wild type, as described more fully in the Summary. The preferred mutant T7 DNA ligase has at least one of the following mutations: E63K (SEQ ID NO:4), K73E (SEQ ID NO:6), K137E (SEQ ID NO:7), K174E (SEQ ID NO:9), E182K (SEQ ID NO:11), K210E (SEQ ID NO: 13), E243K (SEQ ID NO:15), D245R (SEQ ID NO: 17), E268K (SEQ ID NO:19), E272K (SEQ ID NO:21), E289K (SEQ ID NO:23), K295E (SEQ ID NO:25) and D336R (SEQ ID NO:27).

The invention discloses a pennisetum purpureum gene PpbHLH148L and application thereof in improving the cold resistance of plants, and relates to the technical field of gene engineering. The nucleotide sequence of the gene is as shown in SEQ ID NO.5. After the gene is subjected to subcellular localization and expression condition analysis after cold stress, the gene is related to low-temperature tolerance. An overexpression vector of the gene is constructed, Agrobacterium tumefaciens is transferred, then Arabidopsis thaliana infection is carried out, cold tolerance determination is carried out on overexpressed Arabidopsis thaliana, phenotypic difference and physiological index difference between an overexpressed strain after cold treatment and a wild type are observed, and it is found that the gene can significantly improve the low-temperature stress tolerance of Arabidopsis thaliana. And in the low-temperature treatment of the pennisetum alopecuroides transgenic callus, the transgenic callus also shows higher tolerance than the wild callus at low temperature. The pennisetum purpureum gene PpbHLH148L can be used for low-temperature tolerance modification of plants, and has an application prospect in creation of low-temperature-resistant plant materials.

The invention discloses application of a hevea brasiliensis HbCPT8 gene in promoting plant growth and increasing gum yield, and belongs to the field of genetic engineering.The hevea brasiliensis HbCPT8 gene is transferred into kok-saghyz through a plant expression vector to obtain a transgenic kok-saghyz positive plant, and it is found through comparison of the phenotype of the transgenic kok-saghyz and the gum content of the root of the transgenic kok-saghyz that the hevea brasiliensis HbCPT8 gene can be used for promoting plant growth and increasing gum yield. Compared with wild type kok-saghyz, the biomass and the rubber yield of the transgenic kok-saghyz are remarkably improved, which shows that the HbCPT8 gene plays an important role in regulating and controlling rubber synthesis and has important significance in cultivating rubber plant varieties capable of increasing the rubber yield.

The invention belongs to the field of biological agriculture, and particularly relates to a wheat negative regulatory factor TaEDR2 and application thereof. The amino acid sequence of the wheat negative regulatory factor encoding protein provided by the invention is as shown in SEQ ID NO: 1, and the nucleotide sequence of the wheat negative regulatory factor TaEDR2 is as shown in SEQ ID NO: 2. According to the invention, gene editing is carried out in a Fielder wild type material, and a TaEDR2 gene editing plant which shows resistance to stripe rust CYR32 is obtained. It is determined that the TaEDR2 gene plays a negative regulation role in wheat stripe rust resistance, a stripe rust resistant wheat material is created by using the negative regulation factor, and new gene and germplasm resources are provided for wheat stripe rust resistant breeding.

The invention belongs to the technical field of biology, and particularly provides reverse transcriptase UCRT vZ mutants with improved thermal stability, the reverse transcriptase UCRT vZ mutants comprise four single-point mutants, six double mutants and four three mutants, and compared with wild type reverse transcriptase UCRT vZ, the half-life period of the mutants is longer at 65 DEG C; the double-mutant has a better effect, and the half-life period of the double-mutant is about four times that of the wild reverse transcriptase UCRT vZ. The invention also provides a genetically engineered bacterium containing the mutant. The reverse transcriptase UCRT vZ mutant obtained by the construction method provided by the invention has better thermal stability, shows higher thermal stability when being used for reverse transcription of RNA (Ribonucleic Acid) into cDNA (Complementary Deoxyribose Nucleic Acid) at a higher temperature, and has higher application potential.

The invention discloses application of a TaLBD30 protein and a coding gene thereof in regulating and controlling a wheat plant type. The invention belongs to the technical field of biology, and particularly relates to application of TaLBD30 protein and a coding gene thereof to regulation and control of wheat plant types. The protein is any one of the following proteins: A1) a protein with an amino acid sequence as shown in SEQ ID No: 1; a2) a protein which is obtained by substitution and / or deletion and / or addition of amino acid residues on the protein of A1), has 80% or more of identity with the protein of A1) and has the same function as the protein of A1); a3) a fusion protein obtained by connecting a protein tag to the N terminal or / and the C terminal of A1) or A2). TaLBD30 is a transcription factor with transcription inhibition activity, and compared with a wild type, the TaLBD30 gene overexpression shows that after TaLBD30 overexpression, the wheat plant height and ear length are remarkably reduced.

The invention includes a mutant Taq polymerase, which can significantly extend and amplify a target sequence where the extension conditions are time limited to as little as one second. The mutant Taq polymerase, or a biologically active fragment thereof, has one or more substitutions differing from the wild type as shown in Table I.

The invention discloses a rice OsTCP11 gene and application of the rice OsTCP11 gene in rice disease resistance, and the nucleotide sequence of the OsTCP11 gene is as shown in SEQ ID NO.1; the method is realized by knocking out a rice OsTCP11 gene; the knockout is carried out by utilizing a CRISPR / Cas9 gene editing technology; the gene OsTCP11 is knocked out through a CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / associated protein 9) technology, and a knockout mutant ostcp11 is obtained; it is found through rice blast resistance identification that compared with wild type rice, the mutant ostcp11 is more resistant to diseases and small in disease spot area, the number of pyricularia grisea carried by diseased leaves is small, and it is indicated that OsTCP11 negatively regulates the resistance of rice to rice blast. The bacterial leaf blight resistance identification finds that compared with wild rice, the mutant ostcp11 is more resistant to diseases and shorter in disease spots, which indicates that the OsTCP11 negatively regulates the resistance of the rice to the bacterial leaf blight. The method is beneficial to cultivation of rice blast and bacterial leaf disease resistant rice varieties, and provides a basis for later screening of high-resistance and broad-spectrum-resistance rice varieties.

The invention discloses a Chinese orchid flowering regulation gene CsFTL3 as well as an encoding protein and application thereof, and belongs to the field of plant molecular biology. The gene CsFTL3 for regulating and controlling flowering of Chinese cymbidium is separated from flower bud cDNA (complementary deoxyribonucleic acid) of a Chinese cymbidium variety 'Xiaoxiang', and 174 amino acids are encoded by the gene CsFTL3. Through construction of an expression vector of the CsFTL3 gene, overexpression in wild-type arabidopsis thaliana and heterologous overexpression of the CsFTL3 gene in the arabidopsis thaliana, it is found that bolting and flowering periods of the arabidopsis thaliana are significantly delayed, flower stems are significantly thickened and increased, the number of rosette leaves is significantly increased, and the growth period is prolonged, which indicates that the gene has the function of delaying the flowering time of the arabidopsis thaliana. The Chinese orchid flowering regulation and control gene can be used for molecular mechanism research of orchid flowering time, delay Chinese orchid flowering time, promote plant vegetative growth, prolong whole Chinese orchid viewing time and promote Chinese orchid genetic improvement and variety breeding.

The invention discloses a phoebe bournei PbGRF1 gene, and the phoebe bournei PbGRF1 gene has a nucleotide sequence as shown in SEQ ID No: 1, or a sequence which is obtained by deleting, adding and / or substituting one or more nucleotides of the sequence and has an unchanged function. The invention also discloses a protein coded by the phoebe bournei PbGRF1 gene. The invention further discloses a method for promoting plant leaf growth. The invention also discloses an application of the phoebe bournei PbGRF1 gene in promoting the growth of plant leaves. The phoebe bournei PbGRF1 gene is cloned, arabidopsis thaliana is transformed through an agrobacterium tumefaciens mediated inflorescence infection method, a phoebe bournei PbGRF1 gene overexpressed plant is obtained, phenotypic observation shows that the leaf area of the arabidopsis thaliana with the overexpressed phoebe bournei PbGRF1 gene is larger than that of a wild type Col, and meanwhile the fresh weight and the dry weight of the overexpressed plant are larger than that of the wild type Col.

The invention belongs to the technical field of biology, and particularly provides UCRT VI heat-resistant reverse transcriptase mutants with improved thermal stability, the mutants comprise four single-point mutants, six double mutants and four three mutants, and compared with wild-type UCRT VI heat-resistant reverse transcriptase, the mutants have longer half-life period at 65 DEG C; the double mutant has a better effect, and the half-life period of the double mutant is about three times of that of wild type DNA polymerase. The invention also provides a genetically engineered bacterium containing the mutant. The UCRT heat-resistant reverse transcriptase mutant obtained by the construction method provided by the invention has better thermal stability, shows higher thermal stability when being used for synthesizing DNA (Deoxyribonucleic Acid) by reverse transcription at higher temperature, and has higher application potential.

The invention relates to a rice OsABCB7 gene as well as an encoding protein and application thereof. The inventor utilizes a CRISPR / Cas9 gene editing system to obtain two homozygous knockout mutants osabcb7-1 and osabcb7-9 of an OsABCB7 gene, wherein the two homozygous knockout mutants osabcb7-1 and osabcb7-9 take japonica rice Nipponbare as a background and have different editing conditions. Many-year and multi-generation agronomic trait investigation shows that under the normal field planting condition, the grain length, grain width, thousand seed weight, single plant yield and the like in the mature period are all remarkably increased compared with those of a wild type contrast, and the maturing rate has no obvious difference; however, under the continuous high-temperature condition, the maturing rate and the yield of a single plant of the mutant are obviously higher than those of a wild type control. Besides, the mutant rice after salt treatment shows enhanced salt tolerance, so that the OsABCB7 gene can be applied to rice crossbreeding and molecular design breeding to improve the rice yield and enhance the salt tolerance and high temperature resistance of the rice.

The invention discloses an application of a rape BnC05ERF378 gene in improving the waterlogging resistance of a plant. The nucleotide sequence of the rape BnC05ERF378 gene is as shown in SEQ ID NO. 1. According to the invention, a brand new stain-resistant gene BnC05ERF378 is cloned from rape for the first time, and is transferred into arabidopsis thaliana to construct a pure line plant and complete stain resistance identification. Experimental results show that the growth state of the transgenic BnC05ERF378 gene arabidopsis thaliana is remarkably superior to that of a wild type after waterlogging treatment, the transgenic BnC05ERF378 gene arabidopsis thaliana can normally bolt, the survival rate is increased from 12.5% to 37.5%, it is fully proved that the gene can enhance the survivability of the plant in a waterlogging environment by regulating the response mechanism of the plant to waterlogging stress, and the survival rate of the plant is increased from 12.5% to 37.5%. And a key guarantee is provided for stable growth of plants under high-humidity, waterflooding and other adverse conditions. According to the discovery and application of the waterlogging-resistant gene BnC05ERF378 disclosed by the invention, a clear functional gene target is provided for improvement of waterlogging resistance of crops. By constructing an expression vector containing the gene and combining mature technologies such as agrobacterium transformation, the gene can be stably introduced into a target plant, and a pure transgenic plant is obtained.

The invention belongs to the technical field of characterization of target samples, and provides PANX3 biopore protein and application of a mutant and single molecule detection thereof. The PANX3 biopore protein is composed of seven identical subunits, the overall channel is funnel-shaped, and the amino acid sequence is shown as SEQ ID NO: 1. After one or more amino acids of the wild type PANX3 biopore protein are mutated into common amino acids except original amino acids, compared with the wild type PANX3 biopore protein, the mutant can be normally expressed and has stable properties, and the current property and the current signal of the mutant are obviously improved.

The invention discloses a method for targeted creation of a fast-growing new tilapia strain, and belongs to the technical field of fish cultivation. The method specifically comprises the following steps: (1) taking a fragment, as shown in SEQ ID NO.1, on a third exon of a pomc gene as a gRNA target sequence; (2) designing and synthesizing a gRNA primer sequence; (3) amplifying by adopting the gRNA primer sequence and the gRNA plasmid, and then transcribing and purifying to obtain a gRNA fragment; (4) transferring the gRNA fragments into fertilized eggs of tilapia mossambica through gene editing to obtain F0-generation positive fish; and (5) hybridizing the F0-generation positive male fish with the wild-type female fish, screening out F1-generation parent fish, and culturing the F1-generation parent fish to obtain F2-generation tilapia with homozygous deletion mutation. According to the invention, the pomc homozygous knockout line is established in the Nile tilapia mossambica for the first time, the new fast-growing line is a complete gene function deletion model, the cost is lower, the efficiency is higher, and the phenotype can be stably inherited.

The invention relates to a T7 RNA polymerase mutant and application thereof. Specifically, the invention discloses a T7 RNA polymerase mutant, and the polymerase mutant has at least one amino acid mutation at sites corresponding to a wild type T7 RNA polymerase and selected from the following groups: the 44th site, the 45th site, the 46th site, the 172nd site, the 389th site, the 425th site, the 441st site, the 632th site, the 643th site, the 754th site and the 789th site; the serial number of the mutation site is based on the sequence shown in SEQ ID NO: 1, the mutation leads to obvious reduction of a byproduct dsRNA generated in the process of enzyme synthesis of mRNA, and meanwhile, the generation of RNA aggregates is reduced.

The disclosure provides interleukin 21 (IL-21) variants that have (i) decreased or no binding to a human interleukin 21 receptor (IL-21R) as compared to a wild-type human IL-21 polypeptide; or (ii) increased stability as compared to a wild-type human IL-21 polypeptide; or (iii) decreased or no binding to a human interleukin 21 receptor (IL-21R) as compared to a wild-type human IL-21 polypeptide and increased stability as compared to a wild-type human IL-21 polypeptide. The disclosure also provides fusion proteins including an antibody or fragment thereof bound, e.g., covalently linked, to an IL-21 variant.

The invention discloses a gene PtoMYB20 for regulating and controlling salt tolerance and drought resistance of poplars. A nucleotide sequence of the gene comprises a sequence as shown in SEQ ID NO. 1. The gene interference fragment is introduced into a plant to obtain a silent transgenic plant, under the conditions of salt stress, drought stress and salt-drought composite stress, the H2O2 and MDA contents of the silent transgenic plant are lower than those of a wild plant, and the SOD and POD activities of the silent transgenic plant are higher than those of the wild plant, so that the damage degree of the plant is effectively reduced; the method is of great significance in improving salt tolerance and drought resistance of the poplar and relieving damage of adversity stress to forestry yield. The invention also discloses a breeding method of the transgenic poplar with strong salt tolerance and drought resistance, provides a new poplar germplasm which not only improves the stress resistance of the poplar but also does not influence the wood yield, and lays a theoretical and gene resource foundation for the stress-resistant molecular breeding of the poplar.

The invention discloses a method for constructing a Ddx58-R109C point mutation mouse model by using a CRISPR / Cas9 gene editing technology and an application of the Ddx58-R109C point mutation mouse model. The method comprises the following steps: (1) target construction: searching a CDS region of a Ddx58 gene, determining an exon part, and determining a gene mutation site; (2) obtaining a fertilized egg, and injecting Cas9mRNA, gRNA and donor oligo into the fertilized egg of the C57BL / 6J mouse together in vitro through a microinjection technology to obtain an F0-generation hybrid mouse; (3) mating the F0-generation hybrid mouse with the wild type mouse to obtain an F1-generation mouse; (4) the F1-generation heterozygous mice which are identified to be positive are subjected to mating, and homozygous Ddx58-R109C mutant type and wild type mice can be obtained by breeding obtained offspring; (5) phenotypic analysis shows that Ddx58-R109C point mutation mice can have pathological changes of systemic lupus erythematosus and lupus nephritis; the invention can provide an effective research approach and method for researchers to deeply explore the action mechanism of Ddx58-R109C mutation in diseases, and has important significance for explaining the pathogenesis of systemic lupus erythematosus and researching and developing targeted therapeutic drugs.

Provided is the use of overexpression of the auxin transporter PtoPIN1a gene in increasing the biomass of Populus tomentosa. The nucleotide sequence of the auxin transporter PtoPIN1a gene is as shown in SEQ ID No. 1. By means of using a genetic engineering method, wild-type Populus tomentosa is transformed by means of Agrobacterium tumefaciens infection to obtain a tissue-specific PtoPIN1a overexpressing transgenic plant, and subsequently, a plant with a relatively high expression level is selected, by means of positive identification, for phenotype observation. Results show that compared with the wild type in the same period, the PtoPIN1a transgenic plant has an increased plant height, a thicker stem and a significantly increased aboveground biomass. By means of the provided technical cultivation method, a high-biomass high-quality poplar variety is cultivated. The method is of great significance for future molecular breeding to cultivate excellent poplar varieties.

The invention belongs to the technical field of gene engineering, and relates to a cyperus esculentus CeMYB154 gene capable of improving salt tolerance and active oxygen scavenging capacity and application of the cyperus esculentus CeMYB154 gene. The Cyperus esculentus CeMYB154 gene belongs to an MYB transcription factor family, the Cyperus esculentus CeMYB154 gene is screened and transformed into Arabidopsis thaliana through transcriptome sequencing, bioinformatics analysis, gene cloning, overexpression vector construction and recombinant strain overexpression, and analysis finds that under salt stress, compared with a wild type, the root length of a plant with the overexpressed CeMYB154 gene is obviously increased, the growth vigor is better, and the growth rate of the plant with the overexpressed CeMYB154 gene is higher. According to the present invention, the content of MDA and H2O2 is significantly reduced, and the activity of CAT, POD and SOD antioxidant enzymes is significantly enhanced, such that the overexpressed CeMYB154 gene can enhance the salt tolerance and the active oxygen scavenging ability of the plant, and the overexpressed CeMYB154 gene is proved to be the salt tolerance positive regulation factor; the CeMYB154 gene can be used in the field of salt-tolerant stress-resistant breeding of cyperus esculentus, and provides an important gene resource for salt-tolerant molecular breeding.

The present invention belongs to the field of nucleic acid editing, and particularly the technical field of clustered regularly interspaced short palindromic repeats (CRISPRs). Specifically, provided in the present invention is a Cas mutant protein having an improved activity and an improved editing efficiency. Compared with a wild-type parental Cas protein, the Cas mutant protein of the present invention has significantly improved activity, and has a wide application prospect.