17 results about "Homologous sequence" patented technology

The invention provides a separated gene for regulating and controlling the nodulation number of leguminous plants. A coding region sequence of the gene is shown as SEQ ID NO: 1 or SEQ ID NO: 2 or SEQ ID NO: 3 or a homologous sequence of the SEQ ID NO: 1 or SEQ ID NO: 2 or SEQ ID NO: 3. The invention has great theoretical and application values for leguminous plant breeding and related application research thereof.

The present invention relates to a plant comprising a modified Tom2a gene, the wildtype of which comprises SEQ ID No. 1, SEQ ID No. 2, SEQ ID No. 3, SEQ ID No. 4, SEQ ID No. 5, or SEQ ID No. 6, or comprises another homologous sequence having at least 70% sequence identity to SEQ ID No. 1, which modified gene leads to increased resistance to a Tobamovirus. The modification in the Tom2a gene leads to a modified Tom2a protein that has a deletion, a substitution, or an insertion of at least one amino acid when compared to SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10, SEQ ID No. 11, or SEQ ID No. 12, or compared to another homologous sequence having at least 70% sequence similarity to SEQ ID No. 7. The plant is preferably a plant of the family Cucurbitaceae or Solanaceae. The invention further relates to a modified Tom2a gene encoding a Tom2a protein comprising a deletion, a substitution, or an insertion of at least one amino acid when compared to SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10, SEQ ID No. 11, or SEQ ID No. 12, or compared to another homologous sequence having at least 70% sequence similarity to SEQ ID No. 7.

The present invention relates to polynucleotide comprising a lncRNA gene according to SEQ ID No: 2, or according to a sequence having at least 50% sequence identity to SEQ ID No: 2, and an AP2c gene according to SEQ ID No: 3, or according to a sequence having at least 50% sequence identity to SEQ ID No: 3, which polynucleotide confers an increased density of type IV trichomes. The invention further relates to a plant comprising said polynucleotide and a SlASAT4 gene according to SEQ ID No. 21, or comprising a homologous sequence having at least 99.6% sequence identity to SEQ ID No. 21 wherein the presence of the polynucleotide leads to increased density of type IV trichomes and wherein the presence of said SlASAT4 gene leads to an increased ratio of S4:17 / S3:15 acyl sugars produced in glandular trichomes present on said plant when compared to a control plant lacking the polynucleotide and the SlASAT4 gene. Such a plant also was found to be resistant to Bemisia tabaci. The invention also discloses markers for the identification of the polynucleotide and the SlASAT4 gene and their use for the identification and development of Solanum lycopersicum plants. Also methods for identifying, producing and selecting of plants and seed are disclosed.

Provided are methods comprising expressing in a recombinant cell comprising a Cas gene a recombinant error-prone reverse transcriptase (RT) and a recombinant spacer RNA comprising a target sequence for mutagenesis of a DNA sequence in the Cas gene; making a mutagenized cDNA polynucleotide homologous to the DNA sequence in the recombinant cell; expressing a recombinant recombineering system in the recombinant cell; and recombining the mutagenized cDNA with the homologous DNA sequence of the Cas gene in the recombinant cell. Also provided are recombinant cells comprising recombinant coding sequences for a recombinant Cas protein, recombinant error-prone reverse transcriptase (RT), recombinant spacer RNA comprising the target sequence, and recombinant recombineering system.

In a homologous recombination method in which a specific DNA sequence is deleted from a DNA molecule, the DNA molecule is used as the DNA molecule. A DNA molecule is prepared in which a pair of target sequences, which are targets for a site-specific recombinase, are disposed so as to sandwich a specific DNA sequence therebetween and so as to have different orientations from each other, and a pair of homologous sequences, which are DNA sequences having homology, are disposed on the outside of the pair of target sequences disposed so as to sandwich the specific DNA sequence. The site-specific recombinase is caused to act on a DNA molecule, and a specific DNA sequence and a pair of target sequences are deleted from the DNA molecule by homologous recombination between a pair of homologous sequences.

There is provided a homologous recombination method of excising a specific DNA sequence from a DNA molecule, comprising: providing the DNA molecule that has a pair of target sequences as a target of a site-specific recombination enzyme are placed in different directions from each other across the specific DNA sequence and that a pair of homologous sequences as DNA sequences sharing homology with each other are placed on further outside of the pair of target sequences placed across the specific DNA sequence; and making the site-specific recombination enzyme act on the DNA molecule and thereby causing homologous recombination between the pair of homologous sequences, so as to excise the specific DNA sequence along with the pair of target sequences from the DNA molecule.

To provide a method for detecting genetic variation in homologous regions of the genome.SOLUTION: The method combines experimental and analytical approaches to resolve the structure of a genomic region in the genome of a subject whose sequence is highly homologous to one or more other regions of the genome. For example, the genomic region may be a gene, and the highly homologous other region may be a pseudogene. The method involves independent alignment, pairing and analysis of sequence reads from the genomic region and the highly homologous other region to identify genetic variation. Also described herein is a computer-assisted method for such methods.SELECTED DRAWING: Figure 1A

The invention provides a rhodotorula mucilaginosa multi-copy plasmid, rhodotorula mucilaginosa transformation, a target gene efficient overexpression rhodotorula mucilaginosa strain and a construction method thereof, and relates to the technical field of gene engineering. The construction method of the plasmid comprises the following steps: screening out genetics mycin as a resistance marker of wild rhodotorula mucilaginosa; carrying out amplification by taking the plasmid YEp352 as a template and P1-F / R as a primer to obtain a linearized plasmid fragment; the method comprises the following steps: amplifying by taking a DNA (Deoxyribonucleic Acid) sequence of a heredity mycin resistance expression box with a homologous arm as a template and G1-F / R as a primer to obtain a target gene segment with a homologous sequence; and assembling the linearized plasmid fragment and a target gene fragment with a homologous sequence, transforming into an escherichia coli competent cell, and finally identifying a positive bacterial colony. The multi-copy plasmid provided by the invention has multiple cloning sites and selective markers, so that genome modification in rhodotorula mucilaginosa is more flexible and convenient, and a target gene can be stably and efficiently expressed.

The invention discloses a gene editing technology and micro-homologous end connection combined mediated large-fragment DNA efficient fixed-point integration method. According to the method disclosed by the invention, an agrobacterium co-transformation plant which respectively carries a vector containing a gene editing expression cassette and a donor vector containing a micro homologous terminal sequence is utilized, and an exogenous DNA fragment is inserted at a fixed point while a plant target gene is edited; the exogenous DNA fragment and the genome double-strand broken end are more easily subjected to micro homologous sequence mediated end connection, so that the exogenous DNA fragment is integrated into the plant genome at a fixed point. The method provided by the invention has the advantages of accuracy, high efficiency, large integrated fragment and the like, greatly reduces the workloads of genetic transformation, target strain screening, marginal sequence analysis and the like, and has important application value in the fields of molecular medicine agriculture and crop germplasm improvement.

The invention relates to the technical field of DNA cloning, and discloses a universal short-fragment DNA cloning method based on reverse PCR and template-free PCR and application of the universal short-fragment DNA cloning method, and the universal short-fragment DNA cloning method comprises the following cloning steps: taking an annular plasmid as a template, and adopting reverse PCR amplification to obtain a linearized vector; an upstream primer and a downstream primer are designed and synthesized, the 3'ends of the two primers have complementary overlapping regions, the 5 'ends of the two primers contain plasmid homologous sequences, and the two primers are subjected to thermal cycle self-assembly and extension in a PCR reaction system without exogenous template DNA to form a blunt-end double-stranded DNA fragment; and purifying the blunt-end double-stranded DNA fragment and a linearized vector, connecting through homologous recombination, and transforming a host cell to obtain a recombinant plasmid containing a target gene. The universal cloning method for the short-fragment DNA does not depend on any restriction enzyme cutting site, and the vector is universal; meanwhile, an exogenous template DNA is not needed; time and cost are greatly reduced, and the method is suitable for construction of high-throughput sgRNA, epitope tags, promoter elements and the like.

The present invention provides an isolated gene for regulating the nodulation number of leguminous plants, and the coding region sequence of the gene is shown as SEQ ID NO:1 or SEQ ID NO:2 or its homologous sequence. The present invention will have great theoretical and application values for leguminous plant breeding and its related applied research.

The purpose of the present invention is to provide: a double-stranded RNA for inhibiting the expression of both human YAP1 and human WWTR1; and a pharmaceutical composition comprising the same. This double-stranded RNA (dsRNA) for inhibiting the expression of both human YAP1 and human WWTR1 includes a sense strand and an antisense strand. The antisense strand includes a region complementary to a homologous sequence in a human YAP1 transcription product or in a human WWTR1 transcription product. The homologous sequence is a nucleotide sequence formed of 17 nucleotides in the human YAP1 transcription product, or is a nucleotide sequence formed of 17 nucleotides in the human WWTR1 transcription product. When the two nucleotide sequences are aligned, at least 14 nucleotides in consecutive 17 nucleotides are identical. Each of the strands has a length of at least 15 nucleotides. The sense strand and the antisense strand form a double-stranded region.

The application provides a multi-copy plasmid of Rhodotorula mucilaginosa, Rhodotorula mucilaginosa transformation, a Rhodotorula mucilaginosa strain with efficient overexpression of a target gene and a construction method of the Rhodotorula mucilaginosa strain, and relates to the technical field of genetic engineering. The construction method of the plasmid comprises the following steps: screening geneticin as a resistance marker of wild Rhodotorula mucilaginosa; taking plasmid YEp352 as a template and P1-F / R as primers to amplify a linearized plasmid fragment; taking a DNA sequence of a geneticin resistance expression box with a homologous arm as a template and G1-F / R as primers to obtain a target gene fragment with a homologous sequence; assembling the linearized plasmid fragment and the target gene fragment with the homologous sequence, and then transforming into an E. coli competent cell; and finally, positive colony identification is performed. The multi-copy plasmid provided by the application has multiple cloning sites and a selection marker, so that the genome modification in the Rhodotorula mucilaginosa is more flexible and convenient, and the target gene can be stably and efficiently expressed.

The invention provides a separated gene for regulating and controlling the nodulation number of leguminous plants. A coding region sequence of the gene is shown as SEQ ID NO: 1 or SEQ ID NO: 2 or SEQ ID NO: 3 or a homologous sequence of the SEQ ID NO: 1 or SEQ ID NO: 2 or SEQ ID NO: 3. The invention has great theoretical and application values for leguminous plant breeding and related application research thereof.

The present invention relates to a modified WAK gene comprising a deletion and / or a substitution, and / or an insertion when compared to the wild-type sequence comprising SEQ ID No. 1 or compared to a homologous sequence having at least 80% sequence identity to SEQ ID No. 1, wherein said modified WAK gene leads to Corynespora cassiicola resistance in a plant. The function of the encoded WAK protein is reduced, changed or absent. The invention further relates to plants comprising the modified gene and showing resistance to a Corynespora cassiicola.

The invention provides a nucleic acid probe for capturing a target sequence, a design method and application thereof. According to the design method disclosed by the invention, two ends of a probe respectively cover flanking areas of a target sequence. The two ends are connected through a linker, and in addition, a probe with over-high homology with a whole genome is filtered out. According to the method, non-specific binding of homologous sequences can be greatly reduced, data waste caused by repeated sequences is greatly reduced, the capture efficiency of the probe and the detection effect of the depth coefficient are effectively improved, and the designed probe has the detection performance of high specificity, high sensitivity and high positive detection rate.

The invention belongs to the technical field of biology, and provides an artificial chromosome as well as a construction method and application thereof. The artificial chromosome comprises an initial artificial chromosome and a functional gene which are connected; the functional genes comprise a Cas9 gene, a Ura selection marker gene, a gRNA recognition sequence, a homologous sequence 1 and a fragment homologous sequence. The construction method comprises the following steps: connecting a functional gene to an initial artificial chromosome in an enzyme digestion and enzyme ligation manner; transforming a connection product into escherichia coli, and picking positive clones from a chloramphenicol resistant plate; and intracellular assembly of the large DNA fragment: introducing the artificial chromosome and the large DNA fragment to be assembled into a yeast cell, and completing assembly through intracellular homologous recombination. The invention also discloses application of the artificial chromosome in gene cloning and DNA synthesis. According to the artificial chromosome disclosed by the invention, the synthesis efficiency of large-fragment DNA can be improved, the problems of easy breakage and low efficiency in in-vitro operation are avoided, and a tool is provided for subsequent synthesis of a complete genome.