22 results about "Promoter mutation" patented technology

The invention provides a cattle FTO gene promoter mutation site and application thereof, and belongs to the field of biotechnology and genetic engineering, the mutation site is located in an FTO gene promoter region on a cattle genome 18 # chromosome, and the mutation site is specifically characterized in that an A base at a transcriptional start site upstream-33 base position is mutated into a G base (chromosome coordinate: Chr: 22040490); according to the mutation, the expression level of the FTO gene is remarkably improved, and the differentiation capacity of intramuscular fat precursor cells is remarkably enhanced; the FTO gene promoter mutation site provided by the invention not only can increase the gene expression quantity, but also can promote the differentiation of intramuscular fat precursor cells, and provides a solid technical basis for the cultivation of new beef cattle varieties and the increase of intramuscular fat content.

The invention provides a kit for detecting human BRAF gene V600E and TERT gene promoter mutation, and belongs to the technical field of biology. The kit comprises a primer, a probe, a 5 * PCR (Polymerase Chain Reaction)-Enhancer, a 10 * PCR Buffer, dNTP (Deoxyribonucleoside Triphosphate) Mix, MalTaq DNAPlyase, magnesium ions, DEPC (Diethylpyrocarbonate) water, a positive control and a negative control. According to the present invention, the ARMS-PCR detection technology is adopted to detect the specific gene mutation, and the target mutation site is identified by designing the specific primer and the probe, such that the detection specificity and the detection sensitivity are improved; by combining with a Taqman probe technology, the fluorescent signal change in the PCR amplification process can be monitored in real time, so that whether mutation of a BRAF V600E site, a TERT C228T site and a TERT C250T site exists or not is accurately judged; according to the method, pre-amplification of 15 cycles is adopted, then quantitative amplification of 30 cycles is adopted, and when the initial amount of the target DNA is very low, the copy number of the target DNA can be increased, so that the detection accuracy and sensitivity are further improved.

The invention discloses a promoter mutant and an application of the promoter mutant in production of isoleucine. The invention provides a DNA molecule which is any one of the following: A1) a DNA molecule with a nucleotide sequence as shown in SEQ ID No.2; a2) is a variant which is obtained by substitution, deletion or addition of one or more nucleotides at other positions except the 66th nucleotide in the nucleotide sequence as shown in SEQ ID No.2, and the obtained DNA molecule has the same or enhanced promoter activity. Experiments prove that a series of promoter mutants with different strength and regulation characteristics are constructed to replace natural promoters, so that the transcriptional level of a target gene in a key period of fermentation is accurately improved, and the yield of the target gene is increased. The promoter engineering strategy provides a new direction for improving the production performance of industrial microorganisms by optimizing endogenous stress-resistant elements.

The application discloses a ClHAK8 gene promoter variation site related to watermelon salt tolerance and application thereof, and relates to the technical field of biology.The ClHAK8 gene promoter variation site is located at the position of-1534 to-1529 bp in the ClHAK8 gene promoter region; and the nucleotide sequence of the ClHAK8 gene is shown as SEQ ID NO.1.It is first clarified in the application that the presence or absence of the W-box element (TTGACC) at the position of-1534 to-1529 bp in the upstream promoter region of the watermelon ClHAK8 gene is a core molecular switch for regulating watermelon salt tolerance.The application clarifies the molecular mechanism of the variation of the promoter cis-element for affecting the salt stress response of watermelon by regulating the expression of a potassium transport gene, perfects the salt tolerance regulation network of watermelon, and provides a stable and practical detection tool for the molecular marker assisted breeding of salt-tolerant watermelon.

Provided are a promoter mutant that regulates the photosynthetic efficiency of maize leaves, a molecular marker, and the use of the promoter mutant and the molecular marker. The mutant has a nucleotide sequence as shown in SEQ ID NO: 1, and the mutant is obtained by means of inserting an 11.2 kb large fragment into a BSD gene promoter. Further provided is a molecular marker closely linked to the insertion of the 11.2 kb large fragment into the BSD gene promoter. The molecular marker is located at 158317719 bp of the BSD gene promoter, with an allelic variant base of A / G, wherein a maize cross-bred line with the large-fragment insertion has base A at said locus, while a maize cross-bred line without the large-fragment insertion has base G at said locus. Further provided are PCR primers and PCR detection kit for detecting the molecular marker, and the use thereof in terms of improving photosynthetic efficiency traits in plants, breeding new plant varieties, etc.

The invention provides application of a promoter variant in plant breeding, and particularly, an NRAMP5 gene promoter variant is screened for the first time, the variant mutates relative to the 1644-2004bp site of a wild type NRAMP5 gene promoter (SEQ ID NO.1), and the agronomic traits of plants can be remarkably improved, including: (a) the manganese content in plant seeds is not changed; (b) the content of iron and magnesium in plant seeds is increased; and (c) reducing the content of heavy metals such as cadmium, lead and arsenic in the plant seeds.

The present invention provides a bovine FTO gene promoter mutation site and its application, belonging to the fields of biotechnology and genetic engineering. This mutation site is located in the promoter region of the FTO gene on chromosome 18 of the bovine genome, specifically, the A base at the -33 base position upstream of the transcription start site is mutated to the G base (chromosome coordinate: Chr:22040490); this mutation significantly increases the expression level of the FTO gene and significantly enhances the differentiation ability of intramuscular fat precursor cells; the FTO gene promoter mutation site provided by the present invention can not only increase the gene expression level, but also promote the differentiation of intramuscular fat precursor cells, providing a solid technical foundation for the cultivation of new beef cattle breeds and the improvement of intramuscular fat content.

This invention provides a lacUV5(26) mutant and a high-yield strain of lysostaphin, belonging to the field of biotechnology. lacUV5 A promoter mutant, the nucleotide sequence of which is SEQ ID NO.1; an Escherichia coli Nissle 1917 expression system, comprising: expression strain EcN△P, plasmid pSC101- lacUV5( 26 )‑T7RNAP、 Expression vector pET-22b(+); plasmid pSC101- lacUV5( 26 ）‑ T7RNAP The nucleotide sequence is SEQ ID NO.2; the expression strain EcN△P is an EcN with plasmids pMUT1 and pMUT2 knocked out. Using this E. coli Nissle 1917 expression system to express lysostaphin can increase the expression level of lysostaphin in the strain.

The invention relates to a rice OsACC gene promoter mutant resistant to fluazifop-p-butyl or clethodim and application of the rice OsACC gene promoter mutant resistant to fluazifop-p-butyl or clethodim. The rice OsACC gene mutant is a deletion mutant; the sequence of the nucleic acid before mutation is deleted is as shown in SEQ ID No. 1; the length of the sequence of the deleted nucleic acid is 38-97 bp, A in the initiation codon ATG of the OsACC gene is 0 site, the deleted nucleic acid occurs in the 305-209 sites of the upstream of the initiation codon ATG, and at least the basic groups of the 305-268 sites of the upstream of the initiation codon ATG are deleted; the length of the sequence of the deleted nucleic acid is 63-97 bp, the deleted nucleic acid occurs in the 305-209 sites upstream of the initiation codon ATG, and at least the bases at the 305-243 sites upstream of the initiation codon ATG are deleted.

Disclosed are a promoter mutant and use thereof. On the basis of a promoter PsrfA, by mutating core regions and an interval sequence of the two core regions, a PsrfA promoter mutant with improved induced expression activity compared to the wild type is obtained. Meanwhile, also provided is a vector containing the promoter mutant. By using the promoter mutant or the vector, the expression quantity of the expressed protein can be increased. The present invention has wide application prospects in the field of protein expression. Meanwhile, on the basis of the promoter mutant, further provided are a recombinant plasmid and recombinant bacteria with improved NADH pyrophosphatase expression quantity, and the recombinant plasmid and recombinant bacteria can be used in the biocatalysis reaction of reduced nicotinamide mononucleotides, facilitating large-scale efficient production of reduced nicotinamide mononucleotides.

The invention belongs to the technical field of gene engineering, and relates to a promoter mutant, a method for producing L-isoleucine by using the promoter mutant and application of the promoter mutant. The invention provides a promoter mutant, and a DNA (deoxyribonucleic acid) molecule of the promoter mutant is any one of the following: A1) a DNA molecule with a nucleotide sequence as shown in SEQ ID No.2; a2) a DNA molecule which is obtained by carrying out conservative replacement on a plurality of nucleotides except the 152nd site of the SEQ ID No.2 and has the homology of 95% or above and the promoter function. A series of promoter mutants with different strength and regulation characteristics are constructed to replace natural promoters of the promoter mutants, and results show that in high-density fermentation of engineering strains, the strains show higher environmental tolerance and longer stable production period, and the yield and production efficiency of the L-isoleucine are remarkably improved.

The invention discloses a promoter mutant and application thereof in increasing the yield of amino acid, and belongs to the technical field of biology. The promoter mutant at least comprises DNA molecules of which the nucleotide sequences are SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 6. According to the application disclosed by the invention, experiments show that point mutation, namely Paccd5-3Mut and Paccd5-3A201G, is carried out on a Paccd5-3 promoter in a wild type ATCC13869 and corynebacterium glutamicum CGMCC (China General Microbiological Culture Collection Center) No.29950, and both the point mutation and the point mutation are favorable for increasing the yield of the L-glutamic acid.

The present application provides a promoter, an expression cassette comprising the promoter, an expression vector comprising the promoter or the expression cassette, a microorganism comprising the promoter, the expression cassette or the expression vector, and their uses in regulating the expression intensity of a target gene, biocatalysis, or biofermentation. The present application also provides a method for increasing amino acid production.

The invention discloses a promoter mutant of bacillus subtilis and application thereof, and belongs to the technical field of biotechnology and genetic engineering. The invention aims to improve the activity of a promoter of bacillus subtilis. The invention provides a promoter mutant of bacillus subtilis, and the mutant is obtained by taking a promoter gene as shown in SEQ ID NO.1 as a starting gene and carrying out site single-base or multi-base mutation. When the high-activity promoter Psrf mutant is used for mediating alpha-amylase expression, the enzyme activity is greatly improved, and the promoter Psrf mutant has excellent performance in a heterologous expression system, has certain universality and has a wide application prospect in the field of industrial production.

The invention relates to a bacillus subtilis time sequence type promoter and application thereof, and aims to provide a group of time sequence type promoter mutants which have higher transcriptional activity in four different growth periods of bacillus subtilis and keep the dependency characteristic of the growth periods through construction and screening of a promoter random mutation library. The method comprises the following steps: by taking pHT plasmids as a skeleton, placing four dependent promoters in different growth periods at the upstream of a reporter gene, constructing four plasmid vectors of pHT-Phag, PLytR, PspoVG and PmmgA-sfGFP, constructing a random mutation library for the promoters, performing high-throughput preliminary screening on the library by using a flow cytometry screening technology, performing secondary screening by using 96-well plate culture, and performing high-throughput screening on the screened library by using a high-throughput screening technology. Finally, mutants with improved transcriptional activity are successfully obtained, and the dominant mutants are applied to recombinant expression of proteins, so that technical support and reference are provided for enriching expression elements in bacillus subtilis.

The application discloses a Bacillus subtilis promoter mutant and application, and belongs to the field of biotechnology and genetic engineering technology. The application aims to improve the activity of the Bacillus subtilis promoter. The application provides a Bacillus subtilis promoter mutant, wherein the mutant is obtained by performing single-base or multi-base mutation on a promoter gene shown in SEQ ID NO. 1. The high-activity promoter Psrf mutant greatly improves enzyme activity when mediating alpha-amylase expression, and has superior performance in a heterologous expression system, has certain universality, and has a wide application prospect in the industrial production field.

The present invention discloses a method for constructing an artificial small promoter library of Saccharomyces cerevisiae and its application. The present invention provides a method for constructing a mutant library of artificial promoters of Saccharomyces cerevisiae, which includes: randomly mutating all or part of the nucleotides at positions 1-6 from the 3'-end of the nucleotide sequence of the artificial promoter UASF-E-C-core1 of Saccharomyces cerevisiae to obtain a mutant library of artificial promoters of Saccharomyces cerevisiae. The present invention uses the strongest small synthetic promoter obtained by Alper as a scaffold and the green fluorescent protein (GFP) as a reporter gene to construct a kozak library. And it is applied to the metabolic regulation of the endogenous pathway of Saccharomyces cerevisiae to make the genes in the metabolic pathway match the appropriate expression level, avoiding problems such as the inhibition of cell growth caused by the large accumulation of toxic intermediate metabolites or the low yield of target products caused by insufficient expression intensity of key genes.

The invention discloses a constitutive promoter library and application, and relates to a construction method of a bacillus methylophilus promoter library, polynucleotide with promoter activity, a promoter mutant library containing the polynucleotide with the promoter activity, a transcription expression cassette, a recombinant expression vector and a recombinant host cell. And a method for producing a target compound. The invention firstly provides the construction method of the bacillus methylolmethane promoter library, a series of promoter mutants can be obtained through the method, the promoter mutants cover a wide expression intensity range and are wide in universality, and abundant promoter elements are provided for modification of target genes.

The invention discloses a medication auxiliary system for tumor treatment and a pharmaceutical composition. The tumor treatment medication auxiliary system provided by the embodiment of the invention comprises a marker acquisition module used for acquiring a tumor marker of a patient; the medication suggestion module is used for giving corresponding medication suggestions based on the obtained tumor markers; the medication suggestion comprises the following steps: if the tumor marker contains BRAF V600E mutation, TERT promoter mutation and SNP rs2853669TT, recommending an inhibitor which is sensitive to a tumor carrying a triple gene combination of the BRAF V600E mutation, the TERT promoter mutation and the SNP rs2853669TT; the inhibitors comprise a BRAF inhibitor, an MEK inhibitor, an MYC inhibitor, an RAS inhibitor and a PI3K / AKT inhibitor. Expression of BRAF, MEK, MYC, RAS, PI3K / AKT and the like is specifically inhibited, and wide killing on normal cells is avoided. The high specificity improves the selectivity and safety of treatment, and reduces the side effects caused by traditional chemotherapy and radiotherapy.

The invention relates to a rice OsACC gene promoter mutant and application thereof. The rice OsACC gene mutant is a deletion mutant; wherein the deletion mutation occurs in the promoter of the rice OsACC gene, and the sequence of the nucleic acid before the deletion mutation is shown as SEQ ID No.1; the length of the sequence of the deleted nucleic acid is 14 bp to 106 bp, A in an initiation codon ATG of the OsACC gene is 0 site, the deleted nucleic acid occurs in 251 to 146 sites of the upstream of the initiation codon ATG, and at least basic groups of 200 to 187 sites of the upstream of the initiation codon ATG are deleted. The mutant can enable rice to obtain resistance to at least one of herbicide fluazifop-p-butyl, haloxyfop-R-methyl and clethodim.

The invention discloses a Zymomonas mobilis recombinant strain for producing itaconic acid and application of the Zymomonas mobilis recombinant strain. Zymomonas mobilis ZMNP-T7 is taken as a starting strain of the recombinant strain, and cis-aconitic acid decarboxylase gene cadA is integrated at the 1650 site of a genome; a cis-aconitase gene CgacnA is overexpressed, an endogenous isocitrate dehydrogenase gene citC of the recombinant strain is subjected to promoter mutation, and an initiation codon ATG is replaced by GTG or TTG; wherein the nucleotide sequences of the gene cadA, the gene CgacnA and the gene citC are shown as SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 in sequence. Zymomonas mobilis (ZMNP-T7) is taken as a starting strain, production of itaconic acid in the Zymomonas mobilis is realized by introducing a specific exogenous gene cadA, and then the yield of itaconic acid is improved by increasing the copy number of itaconic acid synthesis genes, screening cis-aconitase (Aco) and reducing the activity of isocitrate dehydrogenase.

The application belongs to the technical field of biological medicine, and particularly relates to a SaRNA subgenomic promoter mutant, a mutation method and application thereof. The application obtains a series of promoter mutants with significantly improved expression efficiency by site-directed mutagenesis on the SaRNA subgenomic promoter. Compared with the wild-type promoter, the mutant significantly enhances the expression level of the downstream gene at the cell level and in the animal body. The SaRNA constructed based on the mutant can achieve the same or stronger protein expression effect at a lower dose, and has a wide application prospect in the fields of infectious disease prevention, tumor immunotherapy and protein replacement therapy.