55 results about "Als gene" patented technology

A Candida albicans bloodstream infections cause significant morbidity and mortality in hospitalized patients. Filament formation and adherence to host cells are critical virulence factors of C. albicans. Multiple filamentation regulatory pathways have been discovered, however the downstream effectors of these regulatory pathways remain unknown. The cell surface proteins in the ALS group are downstream effectors of the filamentation regulatory pathway. Particularly, Als1p mediates adherence to endothelial cells in vitro and is required for virulence. The blocking of adherence by the organism is described resulting from the use of a composition and method disclosed herein. Specifically, a pharmaceutical composition comprised of a gene, gene product, or specific antibody to the ALS gene family is administered as a vaccine to generate an immune response capable of blocking adherence of the organism.

The invention discloses a CRISPR / Cpf1 system-mediated homologous recombination method using an RNA transcript as a repair template. The method adopts rice ALS genes as a research object, and constructs a homologous recombination vector. RCR1-RCR2-RDR fragments are transcribed in vitro, a RNP method is used, the RNA transcript is used as the repair template, and homologous recombination repair of target genes is achieved in rice callus. The vector is introduced into the rice callus by a gene gun method, and rice plants with ALS gene fixed-point modification are obtained. The results show that RNA used as the repair template can successfully mediate homologous recombination of the target genes, and the method provides a novel idea for crop breeding, and has strong application potential in agricultural breeding.

The invention discloses a rice ALS (acetolactate synthase) mutant type protein, a mutant type gene and applications of the rice ALS mutant type protein and the mutant type gene. The amino acid sequence of the ALS mutant type protein has following mutation: the 628-poisiton amino acid corresponding to the amino acid sequence of rice ALS has the mutation. The invention further discloses a breeding method of creating herbicide-resistant rice by gene editing. The CRISPR / Cas9 gene editing technology is used for editing the ALS gene for the first time, a T-DNA-knockout new material with the stably inherited herbicide resistant characteristic can be obtained in the T2 generation through offspring screening, and basic agronomic traits of the new material have no obvious change. Compared with breeding based on chemical mutagenesis, hybrid transform breeding and the like, the orderly improved molecular breeding technology based on gene editing has the advantages of being rapid, accurate, efficient and the like, and by means of combination with gene function marked genotype selection, breeding efficiency can be greatly increased and breeding progress is substantially accelerated.

The invention discloses an ALS mutant type gene and further discloses ALS protein coded by the ALS mutant type gene and application of the ALS protein. In a rice ALS gene sequence, 75 nucleotide G changes into nucleotide C, and 339 nucleotide A changes into nucleotide C. The protein is derived from a rice mutant plant resisting an ALS inhibitor herbicide, and compared with a rice wild type ALS sequence, the protein sequence of the ALS protein mutates at the site of Gln25 or Gln113 or Ala237. If green plants express the protein sequence, the green plants can resist an acetolactate synthetase inhibitor herbicide and especially an imidazolone and sulfonylurea herbicide.

The invention discloses KASP marker primers for detecting simultaneous SNP mutations in Ser627Asn and Val643Met of rice ALS genes. In terms of the simultaneously generated SNP mutations in the Ser627Asn and the Val643Met of the rice ALS genes, when the phenotype of the double mutant ALS genes is labeled, the sequence of the double mutant ALS genes is a new unreported gene sequence which can primarily determine the labeled material, and then accurately determine the labeled rice material throughdouble KASP markers in the genes so as to carry out quick, highly accurate and high-throughput identification of anti-acetyl lactate synthase inhibitor herbicides of the rice. When the KASP marker primers are used for molecular marker-assisted breeding of hybrid rice varieties with herbicide resistance, the results of two pairs of KASP markers are validated mutually in order to effectively remove sterile line seedlings obtained via 'Tako' selfing by spraying herbicides.

A Candida albicans bloodstream infections cause significant morbidity and mortality in hospitalized patients. Filament formation and adherence to host cells are critical virulence factors of C. albicans. Multiple filamentation regulatory pathways have been discovered, however the downstream effectors of these regulatory pathways remain unknown. The cell surface proteins in the ALS group are downstream effectors of the filamentation regulatory pathway. Particularly, Als1p mediates adherence to endothelial cells in vitro and is required for virulence. The blocking of adherence by the organism is described resulting from the use of a composition and method disclosed herein. Specifically, a pharmaceutical composition comprised of a gene, gene product, or specific antibody to the ALS gene family is administered as a vaccine to generate an immune response capable of blocking adherence of the organism.

The invention provides a specific PCR (polymerase chain reaction) primer pair for detecting ALS (acetolactate synthetase) inhibitor herbicide-resistant descurainia sophia. The primer pair comprises a primer pair (Seq ID No.1-2) for specific amplification of a B-ALS-1 gene of the descurainia sophia and a primer pair (Seq ID No.3-4) for specific amplification of a B-ALS-2 gene of the descurainia sophia. A PCR detection method adopting the primer pair is excellent in specificity and sensitivity, and a specificity test proves that the two ALS genes of the descurainia sophia can be amplified (figures 1 and 2). As the PCR method is simple, convenient and quick to operate, the sampling detection can be carried out in season based on nucleotide detection, and only 2-4 days are required for a process from sampling to results. The sensitivity and specificity of the method can realize quick identification of suspended ALS inhibitor herbicide-resistant descurainia sophia in the field and confirmation of mutation sites, thus the scientific control of resistant weeds in production practice is guided.

The invention provides an escherichia coli genetically engineered bacterium for the synchronous high yield of L-tryptophan and L-valine, and application thereof. The genetically engineered bacterium is obtained through the steps that on a genome of escherichia coli, trpE (S40F) mutation is introduced while a promoter of tryptophan operon is replaced with a Ptrc promoter; an aroG (S180F) gene controlled by the Ptrc promoter is integrated to a tyrR locus; a serA (H344A, N364A) gene controlled by a Plac promoter is integrated to a yjiV pseudo gene locus; a glnA gene controlled by the Plac promoter is integrated to a ycjV pseudo gene locus; and then a bacillus subtilis alsS gene controlled by the Ptrc promoter is integrated to a yghx pseudo gene locus. Shaking flask fermentation is conducted through a strain to be able to accumulate the L-tryptophan within 22-28 h to 10-14 g / L, meanwhile the accumulation amount of valine reaches 5-7 g / L, and the total acid-producing ability is improved by50% or so compared with that of a tryptophan producing strain; and meanwhile, strain OD600 is different slightly, growth problems are avoided, but the acid-producing ability per strain is obviously improved by 120%, and effective utilization of carbon sources and cells is improved greatly.

The invention discloses a brassica napus mutator gene for resisting imidazolone weedicide and application thereof, relating to the field of plant molecule breeding. Primer pairs are utilized to carry out PCR (Polymerase Chain Reaction) proliferation by using a mutant M9 of brassica napus for resisting imidazolone weedicide as a material to obtain a mutator gene of the mutant with the sequence number of SEQ ID NO.1; and proteins are acquired by translation of the gene with an amino acid sequence of SEQ ID No.2. By applying the conventional breeding method of crossbreeding plants, backcrossing plants and the like, the gene is introduced into other napus varieties or variety series having no resistance to the imidazolone weedicide, so as to improve the durability on the imidazolone weedicide by introducing ALS gene mutation nucleotide sequence target varieties or variety series.

Transformed cells are efficiently selected using a mutant ALS gene having high specificity to PC herbicides. The transformation method comprises the steps of: transforming a host cell with a recombination vector containing a gene of interest and a gene coding for a mutant acetolactate synthase having mutation of glycine corresponds to position 95 of the amino acid sequence of a wild-type acetolactate synthase derived from rice to alanine; culturing the transformed cell obtained in the former step in the presence of a pyrimidinyl carboxy herbicide; and wherein the gene coding for the mutant acetolactate synthase is used as a selection marker.

The invention discloses ALS mutant type genes. The 75th nucleotide of the ALS mutant type genes in an ALS gene sequence of rice mutates from G to nucleotide C, the 339th nucleotide of the ALS mutant type genes in the ALS gene sequence of the rice mutates from A to nucleotide C, and the 710th nucleotide of the ALS mutant type genes in the ALS gene sequence mutates from C to nucleotide T. The invention further discloses application of ALS protein coded by the ALS mutant type genes in the aspect of herbicide resistance. The protein comes from a rice mutant plant resisting ALS inhibitor herbicide. Compared with the rice wild type ALS sequence, the protein sequence of the ALS mutant type genes mutates at Gln25, Gln113 and Ala237 sites. Green plants express the protein sequence, and acetolactate synthetase resistant inhibitor herbicide, particularly imidazolone and sulfonylurea herbicide, can be resisted.

The invention discloses an ALS mutant gene. The nucleotide at the 75th locus of the ALS mutant gene in an ALS gene sequence of rice is changed from G to C, the nucleotide at the 339th locus is changed from A to C, the basic group at the 1099th locus is changed from C to T, and the basic group at the 1880th locus is changed from G to A. The invention further discloses ALS protein coded by the ALS mutant gene and application of the ALS protein. The protein comes from a rice mutant plant resistant to ALS inhibitor herbicide. Compared with a wild ALS sequence of rice, the sequence of the protein is subjected to mutation at the loci of Gln25, Gln113, His367 and Ser627. It is expressed by green plants that the sequence of the protein can resist acetolactate synthetase inhibitor herbicide and particularly imidazolone and sulfonylurea herbicide.

The invention provides a specific polymerase chain reaction (PCR) primer combination for detecting an anti-amyotrophic lateral sclerosis (ALS) inhibitor herbicide beckmannia syzigachne. The combination comprises primer pairs (Seq ID No.1-4) for specific amplification of a beckmannia syzigachne ALS gene. The PCR detection method of adopting the primer combination has excellent specificity and sensitivity. The specificity test proves that the ALS gene of the beckmannia syzigachne can be amplified. The PCR method is simple and fast to operate, sampling detection can be carried out in the season on the basis of detection of nucleotide, the time from sampling to result just is 2-4 days, and rapid identification of a suspected anti-ALS inhibitor beckmannia syzigachne in a field and verification of a mutation site can be achieved by the sensitivity and the specificity, so as to guide scientific management of resistant weeds in production practice.

The invention discloses a wheat ALS mutation gene. The 331th nucleotide of an ALS gene sequence of 6BL chromosome of a wheat B genome is changed into nucleotide A from G. The invention further discloses wheat ALS mutation protein encoded by the wheat ALS mutation gene and application of the wheat ALS mutation protein. The protein is from a wheat mutant plant of an anti-ALS inhibitor herbicide. Compared with a wheat mild ALS sequence, the protein sequence mutates at a Val111 site. A green plant expresses that the mutation protein sequence can be resistant and tolerant to an acetolactate synthase inhibitor herbicide, especially an imidazolone herbicide. After 3mL of imazameth / L water (9-fold recommended concentration) is applied to a wheat seedling with one leaf and one core of the wheat ALS mutation protein, the plant can still normally grow, develop and fruit.

The present invention discloses a system for site-specific modification of ALS gene by a CRISPR-Cas9 system to produce herbicide-resistant rice, and uses thereof. The system for site-specific modification in a plant genome of the present invention comprises a vector for site-specific modification in a plant genome and a donor DNA; wherein the vector for site-specific modification in a plant genome comprises a Cas9 protein expression cassette, gRNA expression cassettes and a donor DNA; the gRNA expression cassettes encode two gRNAs targeting two target sites of a target DNA of a plant of interest, respectively; the target DNA has a fragment to be site-specifically modified which is positioned between the two target site.

The invention relates to a four-primer molecular marker method for identifying ALS (acetolactate synthase) gene mutation generation of rice, and belongs to the field of biotechnology engineering. According to the variation of the rice ALS genes at 1642nd and 1643rd basic groups in a coding region from TG to AT, four specific molecular marker primers are designed and are added into a same PCR (polymerase chain reaction) system; different rice DNA (deoxyribonucleic acid) is subjected to amplification; electrophoresis detection is performed; DNA stripes with different sizes represent homozygotes and heterozygotes with different genotypes of the ALS. The method can be used for fast and accurately identifying the anti-ALS inhibitor type herbicide type gene variation occurring in the rice germplasm resources or breeding groups; in addition, the selection efficiency on the ALS-gene-mutation-containing homozygous genotypes can be improved in the breeding process; the breeding process of the anti-ALS inhibitor type herbicide rice varieties is accelerated.

The invention discloses a wheat ALS mutant gene. According to the mutant gene, the 1180th nucleotide of the ALS gene sequence of the wheat B genome 6BL chromosome is changed from nucleotide G to nucleotide A. The present invention further provides a wheat ALS mutant protein encoded by the wheat ALS mutant gene and an application of the mutant protein. The mutant protein originates from wheat mutant plants capable of resisting ALS inhibition herbicide. Compared with the wheat wild type ALS order, the mutant protein order is mutated in the Ala 394 site. The expression of the protein order through a green plant can resist (endure) acetyl lactate synthase inhibitor herbicide, particularly imidazolidinone herbicide. The 1-leave-1-core wheat seedlings with the wheat ALS mutant protein can still grow, develop and bear fruits normally after an imazameth aqueous solution with a concentration of 3mL imazameth per liter of water (nine times of the recommended concentration) is applied to the seedlings.

A multiplex-polymerase chain reaction (PCR) primer is designed by a sugarcane endogenous gene ALS and an exogenous gene element (including Ubiquitin promoter, NOS promoter, NOS terminator, a Bar gene and a Bt gene). After being optimized, a multiplex-PCR reaction system is established, wherein a template which has 60% Premixtaq<TM> volume, 0.3muM primer concentration and 150ng DNA is added into the reaction system, and the annealing temperature of the reaction system is set to be 56 DEG C; a sugarcane internal standard gene (ALS gene) and five exogenous transgenic elements (including the Ubi promoter, the Bt gene, the Bar gene, the NOS promoter and the NOS terminator) can be detected at the same time by one-time PCR reaction, and the detection limit reaches 1% of the DNA content of the template. The multiplex-PCR reaction system established by the experiment not only is capable of detecting the exogenous gene composition of transgenic sugarcane, but also is capable of effectively detecting other transgenic crops.

The invention discloses a wheat ALS mutant protein. The sequences of an ALS gene of a wheat A genome 6AL chromosome, an ALS gene of a B genome 6BL chromosome and an ALS gene of a wheat D genome 6DL chromosome are mutated. The invention further discloses nucleic acid of the wheat ALS mutant protein and further discloses application of the wheat ALS mutant protein. By expressing the protein sequence, green plants can resist (tolerate) acetolactate synthetase inhibitor type herbicides, especially imidazolone type herbicides. Wheat one-leaf one-core seedlings with the wheat ALS mutant protein still can normally grow, develop and fruit after a herbicide is applied according to the concentration of 1 mL of imazameth per L of water (three times of recommended use concentration).

The invention provides a shepherd's purse drug resistance related molecular marker and a specific PCR (polymerase chain reaction) primer pair for detecting anti-acetolactate synthase inhibitor weedicide shepherd's purse. The PCR primer pair comprises a primer pair for specifically amplifying shepherd's purse JC-ALS gene. The PCR detection method by using the primers has excellent specificity and sensitivity, can implement quick identification of suspected anti-ALS inhibitor shepherd's purse in the field and conformation of mutant sites, thereby instructing the scientific treatment on resistant weed in production practice.

The invention is a divisional application of a patent application named molecular marker related with chemical resistance of shepherd's purse herbs and detection kit of molecular marker in the number of 201410643602.9 and provides a specific DNA (deoxyribonucleic acid) fragment related with chemical resistance of shepherd's purse herbs and a specific PCR (polymerase chain reaction) primer pair of the shepherd's purse herbs resisting ALS (acetolactate synthase) inhibitor herbicides, wherein the specific PCR primer pair comprises a primer pair used for specifically amplifying JC-ALS genes of the shepherd's purse herbs. A PCR detection method adopting the primers has great specificity and sensitivity, can quickly identify the shepherd's purse herbs probably resisting ALS inhibitors in fields and confirm mutation sites and accordingly provides an instruction for scientific treatment of resistant weeds in production practices.

The invention discloses an ALS mutant type gene of japonica rice. The 406th nucleotide on an ALS gene sequence of the japonica rice is mutated into a nucleotide T from G. The invention further discloses an ALS mutant protein encoded by the ALS mutant type gene of the japonica rice and application of the ALS mutant protein. The mutant protein comes from a japonica rice mutant plant capable of resisting ALS inhibitor herbicides. Compared with a wild type ALS sequence, the protein sequence in the invention is mutated at the locus Gly136. Due to green plant expression, the protein sequence can resist (tolerate) the ALS inhibitor herbicides, particularly imidazolone herbicides.

The invention discloses an ALS mutant type gene of japonica rice. The 536th nucleotide on an ALS gene sequence of the japonica rice is mutated into a nucleotide T from C. The invention further discloses an ALS mutant protein encoded by the ALS mutant type gene of the japonica rice and application of the ALS mutant protein. The mutant protein comes from a japonica rice mutant plant capable of resisting ALS inhibitor herbicides. Compared with a wild type ALS sequence, the protein sequence in the invention is mutated at the locus Ala179. Due to green plant expression, the protein sequence can resist (tolerate) the ALS inhibitor herbicides, particularly imidazolone herbicides.

A gene therapy method, which is mediated with a gene vector (including, but not limited to 9 type adeno-associated viruses), directly edits a superoxide dismutase 1 (SOD1, for short) mutant gene based on a CRISPR / Cas9 gene editing system, and treats amyotrophic lateral sclerosis (ALS, for short) in vitro and in transgenic mice, is provided by the invention. A provided recombinant virus has bioactivity, and is hoped to be a candidate virus for ALS gene therapy.

The invention discloses an ALS mutation gene. Nucleotide, at site 75 of an ALS gene sequence of rice, of the ALS mutation gene is changed into nucleotide C from G; and / or nucleotide at side 339 is changed into nucleotide C from A; and / or nucleotide at site 710 is mutated into nucleotide T from C. The invention further discloses an ALS protein coded by the ALS mutation gene and an application thereof. The protein is from a rice mutant plant of an anti-ALS inhibitor herbicide; and compared with a rice wild type ALS sequence, the protein sequence of the ALS mutation gene generates mutation at site Gln25, Gln113 or Ala237. A green plant expresses that the protein sequence can resist acetolactic acid to synthesize the enzyme inhibitor herbicide, especially imidazolone and sulfonylurea herbicides.

The invention discloses a marker primer and a detection kit for detecting SNP mutation of a rape ALS gene and application thereof. The marker primer can directly reflect the resistance of a plant, hasno misidentification due to genetic exchange, and can specifically distinguish and detect the C or T base of the SNP mutation site of the rape ALS1 gene. The marker primer only needs two steps of PCRand fluorescence detection, has low cost, high flux and high specificity, and is particularly suitable for classification screening and identification of different resistance genotypes in the breeding population. A molecular marking method completely eliminates aerosol pollution of a PCR product, environmental pollution of EB and harm of formaldehyde to the human body, and has the remarkable advantages of being simple, reliable, rapid, high in specificity and sensitivity and environmentally friendly. An SNP molecular marker application method is accurate and reliable, easy to operate and suitable for identification of the ALS1 resistance mutation site and assisted selection of breeding.

The invention provides a specific polymerase chain reaction (PCR) primer combination for detecting an anti-amyotrophic lateral sclerosis (ALS) inhibitor herbicide beckmannia syzigachne. The combination comprises primer pairs (Seq ID No.1-4) for specific amplification of a beckmannia syzigachne ALS gene. The PCR detection method of adopting the primer combination has excellent specificity and sensitivity. The specificity test proves that the ALS gene of the beckmannia syzigachne can be amplified. The PCR method is simple and fast to operate, sampling detection can be carried out in the season on the basis of detection of nucleotide, the time from sampling to result just is 2-4 days, and rapid identification of a suspected anti-ALS inhibitor beckmannia syzigachne in a field and verification of a mutation site can be achieved by the sensitivity and the specificity, so as to guide scientific management of resistant weeds in production practice.

The invention discloses a dCAPS (derived cleaved amplified polymorphic sequences) molecular marking method for rapidly screening imidazolinone herbicide resistant rice. Sequencing comparison proves that mutation of amino acid at a 627 locus of each ALS (Acetolactate synthase) gene into N (Asparagine) is a primary cause of resistance of 'Jinjing 818' to imidazolinone herbicides, the ALS gene carrying a resistance locus is named as ALS627N, and the ALS gene without carrying the locus is named as ALS627S. According to DNA (deoxyribonucleic acid) sequence differences, a novel functional markerdC-ALS-627 is developed. The marker can effectively distinguish ALS627N homozygotic type, ALS627S homozygotic type and hemizygous lines. In addition, herbicide spraying tests prove that the ALS627N is a dominant herbicide resistant gene. Rice varieties in different regions are detected by the marker to prove that the marker can be widely used for marker-assisted breeding of the ALS627N gene, canaccelerate culture of new imidazolinone herbicide resistant varieties and has important practical application values.

The invention discloses a method for endowing paddy rice with an herbicide-resistant function through mutation breeding. The method comprises the following steps: (S1) carrying out chemical mutagenesis by virtue of a chemical mutagenic agent containing 4mmol / L of sodium azide, 0.3% of nitrosomethyl urethane and 0.2% of 5-bromouracil; (S2) carrying out CO2 laser radiation induced mutation; (S3) inducing embryonic callusing; (S4) plasma radiation induced mutation: carrying out plasma radiation induced mutation on paddy rice embryonic calluses for 90s-120s, wherein the radiation power is 20W / cm<2>-30W / cm<2>, and the discharging interval is 40mm-50mm; and (S5-S7) screening M1-M3-generation ALS gene mutation type herbicide-resistant paddy rice. According to the method, by carrying out compoundinduced mutation on to-be-modified paddy rice, the mutation efficiency is improved, so that missense mutation occurs in conserved regions in ALS genomes of the to-be-modified, dislocation distributionsites are formed, an enzymatic structure and / or spatial conformation are / or changed, and then the affinity of the paddy rice to an ALS restrained herbicide is reduced.