74 results about "Start codon" patented technology

The present application relates to a kind of deletion mutant nucleic acid and its application in anti-herbicide.The deletion mutation occurs in the promoter of rice OsHPPD Gene, the length of deletion sequence is at least 10 bp, and the A in the start codon ATG of gene is 0, at least the bases located in the upstream of start codon ATG from 2041 to 2032 are deleted. OsHPPD The present application finds that by deleting part of the sequence in the promoter of rice OsHPPD Gene, rice can obtain resistance to HPPD inhibitor herbicides, which is of great application value for ensuring agricultural production safety and improving the efficiency of herbicide use.

The invention discloses application of an upstream non-coding region uORF of an ABA2 gene in regulation and control of crop agronomic traits, in particular to application in improvement of pre-harvest sprouting resistance of rice and cultivation of pre-harvest sprouting resistant rice strains, and belongs to the technical field of gene engineering. The invention finds that uORF in the upstream non-coding region of the crop ABA2 gene can inhibit the translation level of downstream ABA2 protein on the basis of not influencing the transcriptional level, so that the uORF can participate in regulation and control of crop agronomic traits in which ABA participates in regulation and control, such as improvement of pre-harvest sprouting resistance of rice. Therefore, the upstream non-coding region uORF of the crop ABA2 gene can be modified through a hybridization or gene editing technology, so that the purpose of regulating and controlling crop agronomic traits in which ABA participates in regulation and control is achieved, for example, site-directed mutagenesis is performed on an initiation codon of the upstream non-coding region uORF of the rice ABA2 gene, and the pre-harvest sprouting resistance of rice is improved.

The invention discloses a 5 'UTR library, a recombinant expression vector and a genetically engineered bacterium for improving the yield of folamin in pathogenic bacterium Badei and application of the 5' UTR library and the genetically engineered bacterium, and belongs to the technical field of microbial genetic engineering. The method comprises the following steps: designing an SD region based on a 16S rRNA (ribosomal Ribonucleic Acid) 3'end complementary sequence of a Buddei pathogenic bacterium XBD101 serving as an original strain, regulating and controlling the number of bases between the SD sequence and an initiation codon AUG and optimizing an UTR secondary structure to construct a library containing 14 UTR sequences, and replacing three sequences N4A, N7A and N18A with the strongest translation ability in the library to the UTR region of a folamin synthesis key gene fclC, so as to obtain the folamin peptide. And finally, the capacity upgrading of the folamin chassis strain is realized, and an important support is provided for the industrial production of agricultural antibiotics.

The invention discloses an mRNA sequence scoring method and related equipment. The method comprises the following steps: acquiring an mRNA sequence to be scored; extracting a key base sequence from the mRNA sequence, wherein the key base sequence comprises an initiation codon context and a termination codon context; inputting the key base sequence into a target scoring model, and processing to obtain a score of the mRNA sequence in the target index; wherein the target scoring model comprises a multi-modal RNA sequence coding module and a large language model, and the target scoring model is obtained based on unsupervised pre-training of transcription sample data and supervised fine tuning of a score label data set corresponding to the target index. According to the method, a non-coding region and a coding region key region (start / stop codon context) are integrated as input, and a large language model trained based on mass data is used for processing, so that various complex factors such as sequence context information and secondary structures can be comprehensively considered, and the method can be widely applied to the technical field of gene sequence analysis.

The application discloses a method for quantitatively regulating gene translation and application thereof. The method comprises the following steps: sequencing translation efficiency of Kozak sequences and variants thereof before a start codon of a target gene or before an upstream open reading frame of a non-coding region of the target gene, and performing gene editing on the Kozak sequences according to a sequencing result, so as to realize in-situ manipulation of the Kozak sequences and quantitatively regulate gene translation, and the Kozak sequences and the variants thereof are 3-6 bp in length respectively. The application establishes a highly efficient, flexible and widely applicable gene expression regulation method. By using a precise gene editing tool to customize the Kozak sequences of the target gene in-situ, the expression level of the target gene can be quantitatively controlled at the gene translation stage.

A bioengineering method which comprises introducing an inert nucleic acid cassette into a biological entity without introducing or modifying characteristics or traits in the biological entity. The method comprises receiving or providing a sample comprising the biological entity having a nucleic acid sequence; selecting an integration site in the nucleic acid sequence for inserting the inert nucleic acid cassette; designing the inert cassette with optimized primer sequences, optimized probe sequences, optimized stop codons and disrupted start codons, and inserting the inert nucleic acid cassette into the biological entity at the integration site; and validating that no characteristics have been added or modified in the biological entity.

A microbial gene editing scheme automatic design method, system and storage medium; the method comprises: selecting a design object for different types of strains, the design object is bacteria or fungi; after determining the target gene, obtaining the target gene information from the NCBI database; the design scheme of bacteria comprises the following steps: determining the gRNA design region; selecting the optimal gRNA sequence; doing sequence complexity and GC content analysis; the design scheme of fungi uses a frameshift scheme: the gRNA of the frameshift scheme is set at any position not overlapping with other genes after the start codon in the gene; select gRNA sequence in gRNA design region, select the optimal gRNA sequence; design two pairs of four primer sequences for homologous arm identification primer; this scheme can replace the complicated and time-consuming manual scheme, save a lot of time, solve the problem of high cost in the design of the best site and the design of sgRNA sequence scheme of artificial gene knockout.

The invention relates to the technical field of plant breeding, in particular to a mutant of an FZP gene promoter and application of the mutant. Compared with a wild type rice FZP gene promoter, the mutant has any one of the following mutations: (1) 5bp is deleted from 3995bp to 3989bp at the upstream of an initiation codon; and (2) inserting 1bp between 3991bp and 3990bp at the upstream of the initiation codon. The application comprises the following steps: i) increasing the grain density, the spikelet number per ear, the secondary branch number or the yield of the oryza plants; ii) cultivating a transgenic Oryza plant; the FZP gene promoter mutant has the advantages that the FZP gene promoter mutant can be used for regulating and controlling the stem differentiation process of rice, promoting the increase of the grain density of the rice and increasing the yield of the rice, and the FZP gene promoter mutant has an important application value.

This invention relates to the field of gene editing technology, specifically providing a method for creating herbicide-resistant rice by gene editing the promoter region of the OsEPSPS gene. The method involves introducing a WT / -29bp monoallelic deletion mutation into the OsEPSPS gene promoter region. Specifically, with position A of the OsEPSPS gene start codon ATG set to 0, a 29bp deletion is made in one allele located at positions 1122 to 1150 upstream of the start codon ATG, while the other allele remains unchanged. This invention demonstrates that by introducing this monoallelic deletion mutation, rice can acquire resistance to EPSPS inhibitor herbicides. This resistance was identified in the T0 generation plants carrying this mutation and can be stably inherited by their offspring, possessing significant application value for ensuring agricultural production safety and improving herbicide application efficiency.

The application relates to the technical field of wheat kernel type genes, and provides a wheat kernel type gene TaCYT-5A TaCYT-5A The application of the molecular marker is KASP-TaCYT-5A, the marker is a SNP site (C / T) detected at 495818646 bp in a QTL cluster for controlling the wheat kernel type and kernel length traits of a 5A chromosome 484432128-495883712 bp, that is, a 4-hydroxyphenylacetaldehyde oxime monooxygenase gene TaCYT-5A The coding region is 124 bp away from the start codon. The functional KASP molecular marker of the wheat 4-hydroxyphenylacetaldehyde oxime monooxygenase gene TaCYT-5A provided by the application can be used for identifying whether an TaCYT-5A excellent allele exists in a wheat variety / strain, and applied in assisted selection breeding and breeding in combination with other known kernel type related genes.

The invention relates to the technical field of gene editing, and particularly provides a method for creating fluazifop-p-butyl herbicide resistant rice through gene editing of an OsACCase gene promoter region. The method comprises the following steps: constructing a crRNA plasmid library covering an OsACCase gene promoter region on the basis of a CRISPR / LbCas12a system, performing agrobacterium-mediated transformation on rice callus, and performing resistance screening, fluazifop-p-butyl herbicide screening and molecular identification to obtain a fluazifop-p-butyl resistant rice strain. The strain is subjected to-4bp / -5bp double allelic deletion mutation in an upstream 1865-site region of an initiation codon ATG of the OsACCase gene, and a homozygous mutant strain and a double allelic deletion mutant strain can normally grow after being sprayed with 6 times of field recommended dose of fluazifop-p-butyl. The herbicide resistance is realized by editing the gene promoter region, the growth defect of coding region mutation is avoided, the breeding technology is enriched, and weed prevention and control are assisted.

The invention relates to the technical field of gene engineering, in particular to a mutant gene and protein for causing rice flag leaves to become smaller and application of the mutant gene and protein. According to the mutant gene capable of causing the rice flag leaf to become smaller, the nucleotide sequence of the mutant gene is a nucleotide sequence shown as SEQ ID NO.1 inserted into the 153rd site of the nucleotide sequence of a wild rice OsGATA15 gene from an initiation codon ATG. According to the invention, an indica type rice line 93-11 is subjected to < 60 > Co-gamma radiation treatment to obtain a new flag-leaf-reduced mutant ym66, and the ym66 mutant is hybridized with the indica type rice line YHSM to obtain an excellent flag-leaf-reduced restorer line NP27. In the seed production process, the seed production yield is not reduced under the condition that flag leaves of NP27 are not cut, and in addition, the F1 generation shows excellent characters in the aspects of plant type and thousand seed weight.

The application discloses an mRNA sequence scoring method and related equipment, and the method comprises the following steps: acquiring an mRNA sequence to be scored; extracting a key base sequence from the mRNA sequence, wherein the key base sequence comprises a start codon context and a stop codon context; inputting the key base sequence into a target scoring model to obtain a score of the mRNA sequence on a target index; wherein the target scoring model comprises a multi-modal RNA sequence encoding module and a large language model, and the target scoring model is obtained based on unsupervised pre-training of transcription sample data and supervised fine-tuning of a score label data set corresponding to the target index. The application integrates non-coding regions and key regions of coding regions (start / stop codon contexts) as inputs, and processes them by using a large language model trained based on massive data, so that the application can comprehensively consider various complex factors such as sequence context information and secondary structure, and can be widely applied to the technical field of gene sequence analysis.

Provided herein are RNA editing oligonucleotides capable of effecting ADAR-mediated deamination of a target adenosine comprised in a target RNA, wherein the oligonucleotide comprises a sequence that is capable of hybridizing with a region in the target RNA comprising said target adenosine, and wherein the oligonucleotide comprises at least one conformationally restricted nucleotide (CRN) positioned at the 5' and / or at the 3' terminus of the oligonucleotide and / or internally. In particular, the target adenosine may be part of a therapeutic target codon in the MECP2 protein-coding RNA. Provided herein are RNA editing oligonucleotides capable of effecting ADAR-mediated deamination of a target adenosine in a target RNA molecule encoding a target protein, wherein the oligonucleotide comprises a sequence that is capable of hybridizing with a region in the target RNA molecule comprising said target adenosine, and wherein the target adenosine is part of a codon encoding a posttranslational modification site in the target protein. In particular, the target adenosine may be part of a ubiquitination site in the frataxin (FXN) protein. Provided herein are RNA editing oligonucleotides capable of effecting ADAR-mediated deamination of a target adenosine in a target RNA molecule encoding a target protein, wherein the oligonucleotide comprises a sequence that is capable of hybridizing with a region in the target RNA molecule comprising said target adenosine, and wherein the target adenosine is part of a start codon in the 5' untranslated region of the target RNA molecule. In particular, the target adenosine may be part of the start codon of an untranslated ORF (uORF) in the human Sodium-dependent phosphate transporter 2 protein (SLC20A2).

The invention relates to the technical field of gene editing, and particularly provides rice OsEPSPS gene promoter region deletion mutation and application thereof in creation of herbicide-resistant rice, the deletion mutation is that a sequence with the length of 22bp or 28bp is deleted in the OsEPSPS gene promoter region, namely, A in an OsEPSPS gene initiation codon ATG is 0 site, A in the OsEPSPS gene initiation codon ATG is 0 site, A in the OsEPSPS gene initiation codon ATG is 0 site, and A in the OsEPSPS gene initiation codon ATG is 0 site; and deleting the basic groups at sites 2534 to 2555 upstream of the initiation codon ATG or the basic groups at sites 2527 to 2554 upstream of the initiation codon ATG. According to the application disclosed by the invention, the rice can obtain resistance to EPSPS inhibitor herbicides by deleting part of sequences in the promoter of the rice OsEPSPS gene, so that the application has great application values in guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

The invention relates to a rice OsACC gene promoter mutant for resisting ACCase inhibitor herbicides and application of the rice OsACC gene promoter mutant. The rice OsACC gene mutant is a deletion mutant or a deletion and combination insertion mutant; the deletion mutation occurs in the promoter of the rice OsACC gene, and the sequence of the nucleic acid before deletion mutation is as shown in SEQ ID No. 1; the length of the sequence of the deleted nucleic acid is 29 bp to 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 271-243 sites of the upstream of the initiation codon ATG are deleted; the inserted nucleic acid occurs within sites 271 to 243 upstream of the initiation codon ATG. The mutant can enable rice to obtain resistance to at least one of herbicides such as haloxyfop-R-methyl, fluazifop-p-butyl and clethodim.

The invention discloses application of a rice OsME5 gene and a molecular marker thereof in regulation and control of rice dry direct seeding seedling emergence, the nucleotide sequence of the rice OsME5 gene is as shown in SEQ ID No.1, and the coding sequence of OsME5 protein coded by the rice gene OsME5 is as shown in SEQ ID No.2; the method comprises the following steps: knocking out an OsME5 gene by adopting a CRISPR / Cas9 method, and designing a molecular marker by utilizing natural nucleotide polymorphism (SNP) of an OsME5 promoter region SEQ ID No: 3; the OsME5 has a key SNP OsME5-578G / A at 578 base pairs on the upstream of an initiation codon, the seedling emergence ability of the germplasm carrying the OsME5-578G is relatively strong when the germplasm carrying the OsME5-578G is covered with soil and the seedling emergence ability of the germplasm carrying the OsME5-578A is relatively weak when the germplasm carrying the OsME5-578G is covered with The OsME5 gene disclosed by the invention can be applied to the field of agriculture, and a candidate gene is provided for improving the seedling emergence capability of rice soil-covering dry direct seeding.

The present application relates to an oil palm UBQ promoter and application thereof, and belongs to the technical field of biology.The nucleic acid sequence of the promoter is shown as SEQ ID NO.1, the present application firstly aligns and intercepts a sequence of 2000bp upstream of a start codon in an oil palm genome, and synthesizes an oil palm RNA polymerase II type promoter, i.e., an oil palm endogenous ubiquitin promoter EgUBQ.The promoter has transcription activity, and can drive expression of downstream green fluorescent protein NeonGreen.The present application provides an oil palm endogenous UBQ promoter for genetic transformation and gene editing of oil palm.

The invention features lentiviral transfer vectors that include heterologous nucleic acids to be introduced into a cell. The lentiviral transfer vector may be characterized by the following features: (a) including a cytomegalovirus (CMV) promoter; (b) including a polynucleotide encoding a partial gag protein that includes a mutated INS1 inhibitory sequence that reduces restriction of nuclear export of RNA; (c) not including a polynucleotide encoding the INS2, INS3, and INS4 inhibitory sequences of gag; (d) not including an SV40 origin of replication and / or an f1 origin of replication; (e) including a cPPT sequence that contains splice site; (f) including an EF1alpha promoter with intact splice donor and acceptor sites; and (g) including hepatitis B PRE with mutation in start codon of X protein ORF.

The present invention relates to a rice albino leaf and white panicle gene OsAL50 Linked markers with related SNPs and their applications. Rice albino leaf and panicle genes OsAL50 The CDS coding sequence is shown in SEQ ID NO.1, and the protein sequence it encodes is shown in SEQ ID NO.2; and the promoter sequence 3.5 kb upstream of its start codon is shown in SEQ ID NO.3. OsAL50 Molecularly identified SNP-linked markers, including those associated with rice albino leaf and panicle genes OsAL50 Closely linked CAPS-08, CAPS-37 and CAPS-40. OsAL50 Genes can be used in hybrid rice breeding, using OsAL50 The albinism characteristics are displayed to identify and eliminate self-fertile false hybrids at the seedling stage to ensure the purity and quality of the improved varieties.

The application provides a ribosome binding site, a promoter and application thereof. The ribosome binding site is based on an RBS sequence of a trc promoter, a translation initiation region is rationally designed and optimized by using a Salis RBS Calculator, an optimized RBS is obtained, a nucleotide sequence of the RBS is shown in a sequence table SEQ ID NO. 1 or shown in a sequence table SEQ ID NO. 2; a promoter composed of a fixed element upstream of the trc promoter and the optimized RBS is integrated into a valine synthesis pathway gene ygaZH, ilvC, ilvD, ilvE of an Escherichia coli W3110 genome by gene editing technology upstream of an initiation codon ATG, and a valine production strain constructed does not contain a plasmid, has no growth defect, does not need to be induced and has good genetic stability.

An antisense oligonucleotide (ASO) that is capable of binding to a target sequence in a mRNA of a secreted phosphoprotein 1 (SPP1) or osteopontin (OPN) gene, the mRNA having a start codon in an open reading frame encoding the SPP1 or OPN, and the target sequence including the start codon in the open reading frame of the mRNA. Binding of the ASO to the target sequence disrupts translation of the ORF encoding the SPP1 or OPN. Alternatively, an ASO wherein the ASO is capable of binding to a target sequence in a pre-mRNA of a SPP1 or OPN gene. Also double-stranded DNA or RNA molecules capable of binding to a target sequence in a mRNA or pre-mRNA of a SPP1 or OPN gene. Additionally, pharmaceutical compositions and methods of treatments using the ASOs or double-stranded DNA or RNA molecules.

This invention belongs to the fields of genetic engineering and biomedicine, specifically relating to a method for codon optimization of human coagulation factor IX and the recombinant human coagulation factor IX encoding gene and its applications. Based on RiboDecode, this invention optimizes relevant parameters to improve the codons of the target gene, thereby increasing the expression level and activity of the target gene in the host. The optimized FIX gene mRNA molecule exhibits good stability and high translation efficiency; further addition of GCCACC upstream of the start codon further enhances expression efficiency. This invention optimizes transfection-related operations, identifies optimal transfection conditions, and achieves maximum expression levels through transient transfection, effectively improving the yield of recombinant human coagulation factor IX. The results of the examples show that the yield of recombinant human coagulation factor IX can reach 1.58±0.10 mg / L, and the coagulation activity can reach 66.8±0.51%, with significant optimization of all key indicators.

This invention relates to a method for constructing a polycystic kidney disease (PCD) model and its applications. The method involves overexpressing the MYCN gene in a target animal to obtain a PCD model that leads to PCD-related phenotypes. The method includes the following steps: obtaining a first strain of mice with Rosa26 knock-in overexpressing the CAG-LSL-HA tag-MYCN-IRES-BFP-Wpre-polyA gene; obtaining a second strain of mice by inserting Cre-WPRE-polyA into the start codon of the Pax8 gene; and crossing the first and second strains of mice to obtain a MYCN-overexpressing PCD model. This invention employs various experimental methods for validation, including histopathological analysis, immunohistochemical staining, and Western blotting. The model provided by this invention overcomes the limitations of existing in vitro cell models, organoids, and existing animal models in terms of limited phenotypes. The established MYCN-overexpressing PCD animal model exhibits stable disease progression and a short disease cycle, which not only helps to elucidate the disease mechanism but also serves as an ideal platform for drug screening and efficacy evaluation.

This invention belongs to the field of plant molecular biology and genetic breeding technology, specifically relating to a molecular marker related to the malic acid content of apple fruit and its application. The molecular marker is... MdNAC029 The single nucleotide polymorphism site in the gene promoter region, this site is located MdNAC029 The nucleotide sequence of the gene promoter region, starting from the 2693rd base at the 5' end, corresponds to... MdNAC029 At a position 442 bp upstream of the transcription start codon, the polymorphism is either T or C, with the C / C genotype indicating high malic acid content and the T / T genotype indicating low malic acid content. This invention, through molecular biology experiments, confirms that the T allele is the binding site for the transcriptional repressor MdMYC2, and the C allele eliminates the binding of MdMYC2, thereby relieving transcriptional repression and promoting malic acid accumulation. This provides a new molecular marker for the genetic improvement of apple fruit acidity traits, accelerating the breeding process of superior varieties.

A self-circularized RNA structure of the present invention can be expressed in a DNA vector and, at the same time, form circRNA by being circularized through a self-targeting and splicing reaction, wherein the circRNA consists of only a gene of interest. The gene of interest includes an IRES region, an initiation codon and a termination codon, and thus enables the rapid expression of a peptide and a protein.

Disclosed is an mRNA composition containing: an mRNA sequence comprising, in order from 5′ to 3′, a 5′ cap region, a 5′ UTR region, a start codon region, a 3′ UTR region, and a poly (A) tail region; and an antisense oligonucleotide containing a region complementary to the 5′ cap region. The mRNA composition containing the mRNA sequence and the antisense oligonucleotide containing a region complementary to the 5′ cap region of the mRNA sequence may regulate the translation rate of the mRNA, enables selective protein expression based on the type of nucleotide modification and DNA repair mechanism, and may improve the stability of the mRNA against RNA-degrading proteins, thereby improving the stability and efficiency of mRNA vaccines or therapeutics.

The application belongs to the field of aquatic organism breeding, and particularly relates to a rice field eel D2HGDH The application relates to a SNP site combination related to a gene and resistance to aquatic pathogenic viruses and application thereof. D2HGDH The SNP site combination related to the gene and resistance to the rice field eel rhabdovirus is located at the 149th, 365th, 394th, 902nd and 939th base from the start codon of the gene. D2HGDH The detection of the SNP site combination of the gene can screen the individuals resistant to the rhabdovirus of the rice field eel. Therefore, the SNP molecular marker can be used for molecular marker assisted breeding of the rice field eel, accelerates the breeding of the disease-resistant rice field eel, and provides a reference for the disease-resistant breeding of the rice field eel.

The invention discloses an HlyA secretion system for increasing the secretion amount of substrate protein and a construction method and application thereof.The HlyA secretion system comprises a first component, a second component and a third component, the first component comprises an inner membrane transporter HlyB and a membrane interstitial adaptor protein HlyD, and the inner membrane transporter HlyB and the membrane interstitial adaptor protein HlyD are obtained by expressing modified hlyBD operons; according to the modified hlyBD operon, a nucleotide sequence for coding a hexa-polyhistidine tag is inserted behind an hlyD gene initiation codon in the hlyBD operon; and a second component: an outer membrane channel protein TolC derived from a host cell. According to the invention, the HlyD protein is specifically modified, so that the secretion amount of the HlyA secretion system to the substrate protein is obviously increased.

The application provides a 5-aminoacetylpropionic acid-producing strain and a construction method and application thereof. The strain takes Escherichia coli as a chassis, up-regulates the expression amount of a key gene hemA of a 5-aminoacetylpropionic acid synthesis pathway, can improve the carbon flux for synthesizing 5-ALA, and simultaneously inhibits the core gene of a porphyrin precursor synthesis pathway, such as by modifying the RBS and the initiation codon of the hemE gene at the genome level and / or knocking out the hemN gene on the genome, so as to reduce the accumulation of porphyrin intermediates, to realize high-yield 5-ALA, and to effectively avoid the discoloration and deepening of the fermentation broth of the above strain, and to reduce the burden of downstream separation and purification. Further, the 5-aminoacetylpropionic acid dehydrase hemB is weakened, so as to reduce the path of 5-ALA into the porphyrin synthesis pathway, and to improve the 5-ALA yield. Thus, the purpose of improving 5-ALA and avoiding discoloration of the fermentation broth is achieved, and the production cost of 5-ALA is effectively reduced.