15 results about "Start codon" patented technology

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 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 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 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.

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.

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.

A method for designing an mRNA vaccine includes receiving sequence information of a candidate mRNA vaccine, extracting an input sequence including a 25nt primer-binding sequence in a 5′ UTR (Untranslated Region) of the sequence information, a 50nt sequence of the 5′ UTR immediately before a CDS (Coding sequence) region, and a 30nt sequence after a start codon of a coding region, calculating secondary structure information for the input sequence, predicting translation efficiency of the candidate mRNA vaccine by inputting the input sequence and the secondary structure information into a pre-trained deep learning model, and, when the translation efficiency of the candidate mRNA vaccine is equal to or greater than a threshold, generating a final mRNA vaccine sequence by linking a sequence of an antigen protein to the sequence information of the candidate mRNA vaccine.

This invention provides a SNP site, identification kit, identification method, and application for the Jk(a+w) blood type, which triggers immune hemolytic transfusion reactions. The SNP site is the c.613C>T mutation at position 613, starting from the start codon of the SLC14A1 gene coding region. In the proband, the c.613C>T mutation was detected in one allele, while the other allele carried other known inactivating mutations. The allele carrying the c.613C>T mutation resulted in reduced expression of the Kidd protein in the proband's erythrocytes; the other allele, carrying other inactivating mutations, did not express the Kidd protein. The combined effect of these two mutated alleles resulted in the Jk(a+w) phenotype. This provides a genetic variation basis for the establishment of rare blood type banks and the assurance of transfusion compatibility.

This disclosure provides a DNA construct comprising: (a) a start exon encoding a start codon; (b) a first frameshift exon downstream of the start exon, the first frameshift exon including a first receptor splice site, wherein inclusion or exclusion of the first frameshift exon in alternative splicing determines the downstream reading frame of a mature mRNA transcript; and (c) a first specific exon downstream of the first frameshift exon, the first specific exon including a second receptor splice site, a first reporter gene, and a second reporter gene, wherein the first reporter gene is located upstream or downstream of the second reporter gene, and wherein the first reporter gene and the second reporter gene are in different reading frames, wherein exclusion of the first frameshift exon causes the first reporter gene to be in the same reading frame as the start codon, and wherein inclusion of the first frameshift exon causes the second reporter gene to be in the same reading frame as the start codon.

This invention belongs to the field of genetic engineering technology, specifically relating to an optimization method and application of human coagulation factor XI. The optimization method of human coagulation factor XI includes codon optimization using sequence optimization tools, adding a start codon ATG to the 5' end of the human coagulation factor XI sequence, and adding a Kozak sequence before the start codon to obtain optimized human coagulation factor XI; the Kozak sequence is GCCACC. The optimization method of this invention can improve the yield and procoagulant activity of human coagulation factor XI in recombinant expression systems, providing an effective solution to the problems of human coagulation factor XI shortage and insufficient activity.

The application discloses a polynucleotide for improving expression level of a target gene and application thereof. The polynucleotide satisfies the following sequence characteristics: containing a start codon ATG, the start codon ATG being connected with a codon GAT coding aspartic acid, the codon GAT coding aspartic acid being connected with a codon coding serine, wherein the codon coding serine comprises TCA or AGT; the polynucleotide codes at least two negative electric amino acid residues in the first 10 amino acid residues, the negative electric amino acid residues comprising one or both of glutamic acid and aspartic acid; the polynucleotide codes at least one arginine residue in the first 5 amino acid residues. The provided polynucleotide can improve the expression effect of an exogenous protein in Escherichia coli.

This invention discloses a semi-dominant gene for controlling maize plant height. GLK53 This invention relates to and its applications, belonging to the field of maize breeding technology. The invention discovers maize... GLK53 Genetic glk53-1 mutants in GLK53 A G / A point mutation occurs 394 bp downstream of the start codon, resulting in a transcript change from wild-type T2 to mutant T1. The T1 transcript has an extra 9 bases at the end of the second exon, causing the insertion of three amino acids VVR into the Myb DNA-binding domain. This mutation has a semi-dominant inheritance characteristic, increasing maize plant height by 25.84%, the total number of leaves by 3.4, and fresh weight by 15.06%; in F1 hybrids, it still significantly increases plant height by 5.71%–10.07% and fresh weight by 13.26%–15.27%. This invention provides a method based on… glk53-1 The breeding methods and molecular markers for mutant maize can be used to rapidly breed tall maize varieties, which has important application value.